Alkyne antagonists of lysophosphatidic acid receptors

ABSTRACT

Described herein are compounds that are antagonists of lysophosphatidic receptor(s). Also described are pharmaceutical compositions and medicaments that include the compounds described herein, as well as methods of using such antagonists, alone and in combination with other compounds, for treating LPA-dependent or LPA-mediated conditions or diseases.

RELATED APPLICATIONS

This application is filed pursuant to 35 U.S.C. §371 as a United StatesNational Phase Application of International Application No.PCT/US2009/067527, entitled “ALKYNE ANTAGONISTS OF LYSOPHOSPHATIDIC ACIDRECEPTORS” filed Dec. 10, 2009, which claims the benefit of U.S.provisional patent application No. 61/121,862 entitled “ALKYNEANTAGONISTS OF LYSOPHOSPHATIDIC ACID RECEPTORS” filed on Dec. 11, 2008,all of which are incorporated herein by reference in their entirety.

FIELD OF THE INVENTION

Described herein are compounds, methods of making such compounds,pharmaceutical compositions and medicaments comprising such compounds,and methods of using such compounds to treat, prevent or diagnosediseases, disorders or conditions associated with one or more of thelysophosphatidic acid (LPA) receptors.

BACKGROUND OF THE INVENTION

Lysophospholipids are membrane-derived bioactive lipid mediators.Lysophospholipids affect fundamental cellular functions that includeproliferation, differentiation, survival, migration, adhesion, invasion,and morphogensis. These functions influence many biological processesthat include, but are not limited to, neurogensis, angiogenesis, woundhealing, fibrosis, immunity, and carcinogenesis.

Lysophosphatidic acid (LPA) is a lysophospholipid that has been shown toact through sets of specific G protein-coupled receptors (GPCRs) in anautocrine and paracrine fashion. LPA binding to its cognate GPCRs (LPA₁,LPA₂, LPA₃, LPA₄, LPA₅, LPA₆) activates intracellular signaling pathwaysto produce a variety of biological responses. Antagonists of the LPAreceptors find use in the treatment of diseases, disorders or conditionsin which LPA plays a role.

SUMMARY OF THE INVENTION

In one aspect, presented herein are compounds of Formula (I), Formula(II) and Formula (III) that inhibit the physiological activity oflysophosphatidic acid (LPA), and therefore, are useful as agents for thetreatment or prevention of diseases in which inhibition of thephysiological activity of LPA is useful, such as diseases in which anLPA receptor participates, is involved in the etiology or pathology ofthe disease, or is otherwise associated with at least one symptom of thedisease.

In one aspect, the compounds of Formula (I), Formula (II) and Formula(III), are useful for the treatment of fibrosis of organs (liver,kidney, lung, heart and the like), liver diseases (acute hepatatis,chronic hepatitis, liver fibrosis, liver cirrhosis, portal hypertension,regenerative failure, non-alcoholic steatohepatitis (NASH), liverhypofunction, hepatic blood flow disorder, and the like), cellproliferative disease (cancer (solid tumor, solid tumor metastasis,vascular fibroma, myeloma, multiple myeloma, Kaposi's sarcoma, leukemia,chronic lymphocytic leukemia (CLL) and the like) and invasive metastasisof cancer cell, and the like), inflammatory disease (psoriasis,nephropathy, pneumonia and the like), gastrointestinal tract disease(irritable bowel syndrome (IBS), inflammatory bowel disease (IBD),abnormal pancreatic secretion, and the like), renal disease, urinarytract-associated disease (benign prostatic hyperplasia or symptomsassociated with neuropathic bladder disease, spinal cord tumor, herniaof intervertebral disk, spinal canal stenosis, symptoms derived fromdiabetes, lower urinary tract disease (obstruction of lower urinarytract, and the like), inflammatory disease of lower urinary tract,dysuria, frequent urination, and the like), pancreas disease, abnormalangiogenesis-associated disease (arterial obstruction and the like),scleroderma, brain-associated disease (cerebral infarction, cerebralhemorrhage, and the like), neuropathic pain, peripheral neuropathy, andthe like, ocular disease (age-related macular degeneration (AMD),diabetic retinopathy, proliferative vitreoretinopathy (PVR), cicatricialpemphigoid, glaucoma filtration surgery scarring, and the like). In oneaspect, the compounds of Formula (I), Formula (II) and Formula (III),are used in the treatment of fibrotic diseases or conditions.

In one aspect, described herein are compounds of Formula (I), Formula(II) and Formula (III), pharmaceutically acceptable salts,pharmaceutically acceptable solvates, and prodrugs thereof. Compounds ofFormula (I), Formula (II) and Formula (III), are antagonists of at leastone of the LPA receptors selected from LPA₁, LPA₂, LPA₃, LPA₄, LPA₅ andLPA₆. In one embodiment, compounds of Formula (I), Formula (II) andFormula (III) are antagonists of LPA₁. In one embodiment, compounds ofFormula (I), Formula (II) and Formula (III) are antagonists of LPA₁and/or LPA₃. In some embodiments, compounds of Formula (I), Formula (II)and Formula (III) are antagonists of LPA₁ and/or LPA₂. In someembodiments, compounds of Formula (I), Formula (II) and Formula (III)are selective antagonists for one of the LPA receptors relative to theother LPA receptors. In some embodiments, such a selective antagonist isselective for the LPA₁ receptor. In some embodiments, such a selectiveantagonist is selective for the LPA₂ receptor. In some embodiments, sucha selective antagonist is selective for the LPA₃ receptor.

Compounds of Formula (I), Formula (II) and Formula (III) are used in thetreatment of diseases, disorders, or conditions in which activation ofat least one LPA receptor by LPA contributes to the symptomology orprogression of the disease, disorder or condition. These diseases,disorders, or conditions may arise from one or more of a genetic,iatrogenic, immunological, infectious, metabolic, oncological, toxic,surgical, and/or traumatic etiology. In one aspect, the methods,compounds, pharmaceutical compositions, and medicaments described hereincomprise antagonists of LPA receptors. In one aspect, the methods,compounds, pharmaceutical compositions, and medicaments described hereincomprise antagonists of LPA₁.

In one aspect are (3-alkynylphenyl)isoxazole and(4-alkynylphenyl)isoxazole compounds, in which the isoxazole isoptionally substituted in the 3-position and substituted in the4-position with a substituted carbamate, urea or amide moiety, and inwhich the phenyl group is optionally substituted with 2 additionalmoieties, and in which the alkyne is substituted with an optionallinking group that links a carboxylic acid or ester, or achemically/bioisosterically related moiety to the alkyne.

In one aspect, provided herein is a compound of Formula (I)pharmaceutically acceptable salt, pharmaceutically acceptable solvate,or prodrug thereof:

-   -   wherein,    -   R¹ is —CO₂H, —CO₂R^(A), —C(═O)NHSO₂R¹⁰, —C(═O)N(R⁹)₂,        —C(═O)NH—OH, —C(═O)NH—CN, tetrazolyl, —OH, —OR⁹, substituted or        unsubstituted monocyclic heteroaryl, or substituted or        unsubstituted monocyclic heterocycloalkyl;    -   R^(A) is H or C₁-C₆alkyl;    -   L¹ is absent, a substituted or unsubstituted C₁-C₆alkylene, a        substituted or unsubstituted C₁-C₆fluoroalkylene, a substituted        or unsubstituted C₁-C₆heteroalkylene, a substituted or        unsubstituted C₃-C₆cycloalkylene, a substituted or unsubstituted        C₁-C₆heterocycloalkylene, a substituted or unsubstituted        phenylene, a substituted or unsubstituted heteroarylene, or        -L²-B-L³-;        -   L² is absent, a substituted or unsubstituted C₁-C₆alkylene,            a substituted or unsubstituted C₁-C₆fluoroalkylene, or a            substituted or unsubstituted C₁-C₆heteroalkylene;        -   B is a substituted or unsubstituted C₃-C₆cycloalkylene, a            substituted or unsubstituted C₂-C₆heterocycloalkylene, a            substituted or unsubstituted phenylene, or a substituted or            unsubstituted heteroarylene;        -   L³ is absent, a substituted or unsubstituted C₁-C₆alkylene,            a substituted or unsubstituted C₁-C₆fluoroalkylene, or a            substituted or unsubstituted C₁-C₆heteroalkylene;    -   R³ is H, C₁-C₄alkyl, or C₁-C₄fluoroalkyl;    -   R⁴ is —NR⁷C(═O)OCH(R⁸)—CY, or —NR⁷C(═O)O—CY;        -   R⁷ is H or C₁-C₄alkyl;        -   R⁸ is H, C₁-C₄alkyl, or C₁-C₄fluoroalkyl;        -   CY is a substituted or unsubstituted C₃-C₁₀cycloalkyl, a            substituted or unsubstituted C₂-C₁₀heterocycloalkyl, a            substituted or unsubstituted phenyl, a substituted or            unsubstituted napthyl, or a substituted or unsubstituted            monocyclic or bicyclic heteroaryl; or        -   R⁸ and CY are taken together with the carbon atom to which            they are attached to form a substituted or unsubstituted            C₃-C₁₀carbocycle or a substituted or unsubstituted            C₂-C₁₀heterocycle;

R⁵ and R⁶ are each independently selected from H, halogen, —CN, —NO₂,—OH, —OR¹⁰, —S(═O)₂R¹⁰, —N(R⁹)S(═O)₂R¹⁰, —S(═O)₂N(R⁹)₂, —C(═O)R¹⁰,—OC(═O)R¹⁰, —CO₂R⁹, —OCO₂R¹⁰, —N(R⁹)₂, —C(═O)N(R⁹)₂, —OC(═O)N(R⁹)₂,—NR⁹C(═O)N(R⁹)₂, —NR⁹C(═O)R¹⁰, —NR⁹C(═O)OR¹⁰, C₁-C₄alkyl,C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, C₁-C₄alkoxy, and C₁-C₄heteroalkyl;

-   -   each R⁹ is independently selected from H, C₁-C₆alkyl,        C₁-C₆heteroalkyl, C₁-C₆fluoroalkyl, a substituted or        unsubstituted C₃-C₁₀cycloalkyl, a substituted or unsubstituted        C₂-C₁₀heterocycloalkyl, a substituted or unsubstituted aryl, a        substituted or unsubstituted heteroaryl, substituted or        unsubstituted —C₁-C₄alkylene-(substituted or unsubstituted        C₃-C₁₀cycloalkyl), —C₁-C₄alkylene-(substituted or unsubstituted        heterocycloalkyl), —C₁-C₄alkylene-(substituted or unsubstituted        aryl), or —C₁-C₄alkylene-(substituted or unsubstituted        heteroaryl); or    -   two R⁹ groups attached to the same N atom are taken together        with the N atom to which they are attached to form a substituted        or unsubstituted heterocycle;    -   R¹⁰ is selected from C₁-C₆alkyl, C₁-C₆heteroalkyl,        C₁-C₆fluoroalkyl, a substituted or unsubstituted cycloalkyl, a        substituted or unsubstituted heterocycloalkyl, a substituted or        unsubstituted aryl, a substituted or unsubstituted heteroaryl,        —C₁-C₄alkylene-(substituted or unsubstituted cycloalkyl),        —C₁-C₄alkylene-(substituted or unsubstituted heterocycloalkyl),        —C₁-C₄alkylene-(substituted or unsubstituted aryl), and        —C₁-C₄alkylene-(substituted or unsubstituted heteroaryl).

For any and all of the embodiments, substituents are selected from amongfrom a subset of the listed alternatives. For example, in someembodiments, R³ is H, C₁-C₄alkyl, or C₁-C₄fluoroalkyl. In otherembodiments, R³ is C₁-C₄alkyl, or C₁-C₄fluoroalkyl. In yet otherembodiments, R³ is C₁-C₄alkyl. In some embodiments, R³ is —CH₃ or—CH₂CH₃. In some embodiments, R³ is —CH₃, —CF₃, —CH₂CH₃, or —CHCH₂. Insome embodiments, R³ is —CH₃.

In some embodiments, R¹ is —CO₂H, —CO₂R^(A), —C(═O)NHSO₂R¹⁰,—C(═O)N(R⁹)₂, tetrazolyl, —OH, substituted or unsubstituted monocyclicheteroaryl, or substituted or unsubstituted monocyclic heterocycloalkyl.In some embodiments, R¹ is —CO₂H, —CO₂R^(A), —C(═O)NHSO₂R¹⁰,—C(═O)N(R⁹)₂, tetrazolyl, —OH, substituted or unsubstituted monocyclicC₁-C₅heteroaryl, or substituted or unsubstituted monocyclicC₁-C₅heterocycloalkyl. In some embodiments, R¹ is —CO₂H, —CO₂R^(A),—C(═O)NHSO₂R¹⁰, —C(═O)N(R⁹)₂, tetrazolyl, —OH, substituted orunsubstituted monocyclic C₁-C₅heteroaryl containing 1-4 N atoms, orsubstituted or unsubstituted monocyclic C₁-C₅heterocycloalkyl containing1-4 N atoms.

In some embodiments, R⁵ and R⁶ are each independently selected from H,halogen, —CN, —NO₂, —OH, —NH₂, —NH(C₁-C₆alkyl), —N(C₁-C₆alkyl)₂,C₁-C₄alkyl, C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, C₁-C₄alkoxy, andC₁-C₄heteroalkyl. In other embodiments, R⁵ and R⁶ are each independentlyselected from H, halogen, —OH, —NH₂, —NH(CH₃), —N(CH₃)₂, —CH₃, —CF₃,—OCF₃, and —OCH₃. In other embodiments, R⁵ and R⁶ are each independentlyselected from H, halogen, —OH, —CH₃, —CF₃, —OCF₃, and —OCH₃. In yetother embodiments, R⁵ and R⁶ are each independently selected from H,halogen, —CH₃, and —CF₃. In some embodiments, R⁵ and R⁶ are each H.

In some embodiments, R¹ is —CO₂H, —CO₂R^(A), or a substituted orunsubstituted monocyclic heteroaryl; R³ is C₁-C₄alkyl; R⁴ is—NR⁷C(═O)OCH(R⁸)—CY; R⁷ is H; R^(A) is H, —CH₃, or —CH₂CH₃.

In some embodiments, R⁵ and R⁶ are each independently selected from H,halogen, —OH, —CH₃, —CF₃, —OCF₃, and —OCH₃; CY is a substituted orunsubstituted C₃-C₆cycloalkyl, a substituted or unsubstituted phenyl, ora substituted or unsubstituted monocyclic heteroaryl containing 0-4 Natoms.

In some embodiments, R¹ is —CO₂H, —CO₂R^(A), or a substituted orunsubstituted monocyclic C₁-C₅heteroaryl.

In some embodiments, L¹ is absent, C₁-C₆alkylene, C₁-C₆fluoroalkylene,C₁-C₆heteroalkylene, a substituted or unsubstituted C₃-C₆cycloalkylene,a substituted or unsubstituted C₁-C₆heterocycloalkylene, a substitutedor unsubstituted phenylene, a substituted or unsubstitutedheteroarylene, or -L²-B-L³-. In some embodiments, L¹ is absent,C₁-C₆alkylene, or -L²-B-L³-.

In some embodiments, R³ is —CH₃ or —CH₂CH₃; L¹ is absent, C₁-C₆alkylene,C₁-C₆fluoroalkylene, C₁-C₆heteroalkylene, a substituted or unsubstitutedC₃-C₆cycloalkylene, a substituted or unsubstituted phenylene, asubstituted or unsubstituted monocyclic heteroarylene containing 0-4 Natoms, or -L²-B-L³-.

In some embodiments, L² is absent, C₁-C₆alkylene, C₁-C₆fluoroalkylene,or C₁-C₆heteroalkylene; B is a substituted or unsubstitutedC₃-C₆cycloalkylene, a substituted or unsubstitutedC₂-C₆heterocycloalkylene, a substituted or unsubstituted phenylene, or asubstituted or unsubstituted monocyclic heteroarylene; L³ is absent,C₁-C₆alkylene, C₁-C₆fluoroalkylene, or C₁-C₆heteroalkylene.

In some embodiments, B is a substituted or unsubstituted phenylene, or asubstituted or unsubstituted monocyclic heteroarylene containing 0-4 Natoms.

In some embodiments, L¹ is a C₁-C₆alkylene, C₁-C₆fluoroalkylene, orC₁-C₆heteroalkylene; R⁴ is

R⁸ is —CH₃ or —CF₃; CY is a substituted or unsubstituted phenyl. In someembodiments, L¹ is a C₁-C₆alkylene.

In some embodiments, R¹ is —CO₂H, —CO₂R^(A), or a substituted orunsubstituted monocyclic heteroaryl containing 1-4 N atoms; In someembodiments, L¹ is -L²-B-L³-; R⁴ is

R⁸ is —CH₃ or —CF₃; CY is a substituted or unsubstituted phenyl.

In some embodiments, R⁴ is

In some embodiments, R⁴ is

In some embodiments, if CY is substituted, then CY is substituted with 1or 2 R^(C), where each R^(C) is independently halogen, —OH, C₁-C₄alkyl,C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, C₁-C₄alkoxy, or C₁-C₄heteroalkyl.In some embodiments, if CY is substituted, then CY is substituted with 1or 2 R^(C), where each R^(C) is independently halogen, —OH, C₁-C₄alkyl,C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, or C₁-C₄alkoxy. In someembodiments, if CY is substituted, then CY is substituted with 1 or 2R^(C), where each R^(C) is independently halogen, —OH, —CH₃, —CF₃,—OCF₃, —OCH₃. In some embodiments, if CY is substituted, then CY issubstituted with 1 or 2 R^(C), where each R^(C) is independentlyhalogen. In some embodiments, if CY is substituted, then CY issubstituted with 1 R^(C).

In some embodiments, R¹ is —CO₂H or —CO₂R^(A). In some embodiments, R¹is —CO₂H.

In some embodiments, R¹ is a substituted or unsubstituted monocyclicheteroaryl containing 1-4 N atoms. In some embodiments, R¹ is asubstituted or unsubstituted 5-membered or 6-membered monocyclicheteroaryl containing 1-4 N atoms. In some embodiments, R¹ is asubstituted or unsubstituted 5-membered monocyclic heteroaryl containing1-4 N atoms. In some embodiments, R¹ is a substituted or unsubstituted6-membered monocyclic heteroaryl containing 1-4 N atoms.

In some embodiments, R¹ is —CO₂H, —CO₂R^(A), or a substituted orunsubstituted monocyclic heteroaryl containing 1-4 N atoms; L² is abond, C₁-C₆alkylene, C₁-C₆fluoroalkylene, —C₁-C₆alkylene-O—,—C₁-C₆alkylene-S—, —C₁-C₆alkylene-S(═O)—, —C₁-C₆alkylene-SO₂—, or—C₁-C₄alkylene-NR²—; B is a substituted or unsubstituted phenylene, or asubstituted or unsubstituted monocyclic heteroarylene containing 1-4 Natoms; L³ is a bond, C₁-C₆alkylene, C₁-C₆fluoroalkylene,—O—C₁-C₆alkylene-, —S—C₁-C₆alkylene-, —S(═O)—C₁-C₆alkylene-,—SO₂—C₁-C₆alkylene-, or —NR²—C₁-C₄alkylene; R² is H, C₁-C₂alkyl, orC₁-C₂fluoroalkyl.

In some embodiments, L² is a bond, C₁-C₆alkylene, C₁-C₆fluoroalkylene,—C₁-C₆alkylene-O—, —C₁-C₆alkylene-S—, —C₁-C₆alkylene-S(═O)—, or—C₁-C₆alkylene-SO₂—. In some embodiments, L² is a bond, C₁-C₆alkylene,C₁-C₆fluoroalkylene, -C₁-C₆alkylene-O—, or —C₁-C₆alkylene-S—. In someembodiments, L² is a bond, C₁-C₄alkylene, C₁-C₄fluoroalkylene,—C₁-C₄alkylene-O—, or —C₁-C₄alkylene-S—. In some embodiments, L² is abond, C₁-C₂alkylene, —C₁-C₂alkylene-O—, or —C₁-C₂alkylene-S—. In someembodiments, L² is a bond or C₁-C₆alkylene. In some embodiments, L² is abond or C₁-C₄alkylene.

In some embodiments, L³ is a bond, C₁-C₆alkylene, C₁-C₆fluoroalkylene,—O—C₁-C₆alkylene-, —S—C₁-C₆alkylene-, —S(═O)—C₁-C₆alkylene-, or—SO₂—C₁-C₆alkylene. In some embodiments, L³ is a bond, C₁-C₆alkylene,C₁-C₆fluoroalkylene, —O—C₁-C₆alkylene-, or —S—C₁-C₆alkylene-. In someembodiments, L³ is a bond, C₁-C₄alkylene, C₁-C₄fluoroalkylene,—O—C₁-C₄alkylene-, or —S—C₁-C₄alkylene-. In some embodiments, L³ is abond, C₁-C₂alkylene, —O—C₁-C₂alkylene-, or —S—C₁-C₂alkylene-. In someembodiments, L³ is a bond or C₁-C₆alkylene. In some embodiments, L³ is abond or C₁-C₄alkylene.

In some embodiments, the compound of Formula (I) has the structure ofFormula (II) or Formula (III):

In some embodiments, the compound of Formula (I) has the structure ofFormula (II).

In some embodiments, the compound of Formula (I) has the structure ofFormula (III).

Any combination of the groups described above for the various variablesis contemplated herein. Throughout the specification, groups andsubstituents thereof are chosen by one skilled in the field to providestable moieties and compounds.

In one aspect, provided is a compound presented in Table 1, Table 2and/or Table 3.

Compounds of Formula (I), Formula (II) and Formula (III) are antagonistsof a LPA receptor.

In some embodiments, the compound of Formula (I), Formula (II) orFormula (III) is an antagonist of LPA₁.

In some embodiments, the compound of Formula (I), Formula (II) orFormula (III) is an antagonist of LPA₂.

In some embodiments, the compound of Formula (I), Formula (II) orFormula (III) is an antagonist of LPA₃.

In some embodiments, presented herein are compounds selected from activemetabolites, tautomers, pharmaceutically acceptable solvates,pharmaceutically acceptable salts or prodrugs of a compound of Formula(I), Formula (II) or Formula (III).

In some embodiments, provided is a pharmaceutical composition comprisinga therapeutically effective amount of a compound of Formula (I), Formula(II) or Formula (III). In some embodiments, the pharmaceuticalcomposition also contains at least one pharmaceutically acceptableinactive ingredient.

In some embodiments, provided is a pharmaceutical composition comprisinga therapeutically effective amount of a compound of Formula (I), Formula(II) or Formula (III), or a pharmaceutically acceptable salt thereof,and at least one pharmaceutically acceptable inactive ingredient. In oneaspect, the pharmaceutical composition is formulated for intravenousinjection, subcutaneous injection, oral administration, inhalation,nasal administration, topical administration, ophthalmic administrationor otic administration. In some embodiments, the pharmaceuticalcomposition is a tablet, a pill, a capsule, a liquid, an inhalant, anasal spray solution, a suppository, a suspension, a gel, a colloid, adispersion, a suspension, a solution, an emulsion, an ointment, alotion, an eye drop or an ear drop.

In some embodiments, the pharmaceutical composition further comprisesone or more additional therapeutically active agents selected from:corticosteroids, immunosuppresants, analgesics, anti-cancer agent,anti-inflammatories, chemokine receptor antagonists, bronchodilators,leukotriene receptor antagonists, leukotriene formation inhibitors,monoacylglycerol kinase inhibitors, phospholipase A₁ inhibitors,phospholipase A₂ inhibitors, and lysophospholipase D (lysoPLD)inhibitors, autotaxin inhibitors, decongestants, antihistamines,mucolytics, anticholinergics, antitussives, expectorants, and β-2agonists.

In some embodiments, provided is a method comprising administering acompound of Formula (I), Formula (II) or Formula (III) to a human with aLPA-dependent or LPA-mediated disease or condition. In some embodiments,the human is already being administered one or more additionaltherapeutically active agents other than a compound of Formula (I),Formula (II) or Formula (III). In some embodiments, the method furthercomprises administering one or more additional therapeutically activeagents other than a compound of Formula (I), Formula (II) or Formula(III).

In some embodiments, the one or more additional therapeutically activeagents other than a compound of Formula (I), Formula (II) or Formula(III) are selected from: corticosteroids, immunosuppresants, analgesics,anti-cancer agent, anti-inflammatories, chemokine receptor antagonists,bronchodilators, leukotriene receptor antagonists, leukotriene formationinhibitors, monoacylglycerol kinase inhibitors, phospholipase A₁inhibitors, phospholipase A₂ inhibitors, and lysophospholipase D(lysoPLD) inhibitors, autotaxin inhibitors, decongestants,antihistamines, mucolytics, anticholinergics, antitussives,expectorants, and β-2 agonists.

Pharmaceutical formulations described herein are administerable to asubject in a variety of ways by multiple administration routes,including but not limited to, oral, parenteral (e.g., intravenous,subcutaneous, intramuscular), intranasal, buccal, topical or transdermaladministration routes. The pharmaceutical formulations described hereininclude, but are not limited to, aqueous liquid dispersions,self-emulsifying dispersions, solid solutions, liposomal dispersions,aerosols, solid dosage forms, powders, immediate release formulations,controlled release formulations, fast melt formulations, tablets,capsules, pills, delayed release formulations, extended releaseformulations, pulsatile release formulations, multiparticulateformulations, and mixed immediate and controlled release formulations.

In some embodiments, the compound of Formula (I), Formula (II) orFormula (III) is administered orally.

In some embodiments, the compound of Formula (I), Formula (II) orFormula (III) is administered topically. In such embodiments, thecompound of Formula (I), Formula (II) or Formula (III) is formulatedinto a variety of topically administrable compositions, such assolutions, suspensions, lotions, gels, pastes, shampoos, scrubs, rubs,smears, medicated sticks, medicated bandages, balms, creams orointments. Such pharmaceutical compounds can contain solubilizers,stabilizers, tonicity enhancing agents, buffers and preservatives. Inone aspect, the compound of Formula (I), Formula (II) or Formula (III)is administered topically to the skin.

In another aspect, the compound of Formula (I), Formula (II) or Formula(III) is administered by inhalation. In one embodiment, the compound ofFormula (I), Formula (II) or Formula (III) is administered by inhalationthat directly targets the pulmonary system.

In another aspect, the compound of Formula (I), Formula (II) or Formula(III) is formulated for intranasal administration. Such formulationsinclude nasal sprays, nasal mists, and the like.

In another aspect, the compound of Formula (I), Formula (II) or Formula(III) is formulated as eye drops.

In another aspect is the use of a compound of Formula (I), Formula (II)or Formula (III) in the manufacture of a medicament for treating adisease, disorder or conditions in which the activity of at least oneLPA receptor contributes to the pathology and/or symptoms of the diseaseor condition. In one embodiment of this aspect, the LPA is selected fromLPA₁, LPA₂, LPA₃, LPA₄, LPA₅ and LPA₆. In one aspect, the LPA receptoris LPA₁. In one aspect, the disease or condition is any of the diseasesor conditions specified herein.

In any of the aforementioned aspects are further embodiments in which:(a) the effective amount of the compound of Formula (I), Formula (II) orFormula (III) is systemically administered to the mammal; and/or (b) theeffective amount of the compound is administered orally to the mammal;and/or (c) the effective amount of the compound is intravenouslyadministered to the mammal; and/or (d) the effective amount of thecompound is administered by inhalation; and/or (e) the effective amountof the compound is administered by nasal administration; or and/or (f)the effective amount of the compound is administered by injection to themammal; and/or (g) the effective amount of the compound is administeredtopically to the mammal; and/or (h) the effective amount of the compoundis administered by ophthalmic administration; and/or (i) the effectiveamount of the compound is administered rectally to the mammal; and/or(j) the effective amount is administered non-systemically or locally tothe mammal.

In any of the aforementioned aspects are further embodiments comprisingsingle administrations of the effective amount of the compound,including further embodiments in which (i) the compound is administeredonce; (ii) the compound is administered to the mammal multiple timesover the span of one day; (iii) continually; or (iv) continuously.

In any of the aforementioned aspects are further embodiments comprisingmultiple administrations of the effective amount of the compound,including further embodiments in which (i) the compound is administeredcontinuously or intermittently: as in a a single dose; (ii) the timebetween multiple administrations is every 6 hours; (iii) the compound isadministered to the mammal every 8 hours; (iv) the compound isadministered to the mammal every 12 hours; (v) the compound isadministered to the mammal every 24 hours. In further or alternativeembodiments, the method comprises a drug holiday, wherein theadministration of the compound is temporarily suspended or the dose ofthe compound being administered is temporarily reduced; at the end ofthe drug holiday, dosing of the compound is resumed. In one embodiment,the length of the drug holiday varies from 2 days to 1 year.

Also provided is a method of inhibiting the physiological activity ofLPA in a mammal comprising administering a therapeutically effectiveamount of a compound of Formula (I), Formula (II) or Formula (III) or apharmaceutically acceptable salt thereof to the mammal in need thereof.

In one aspect, provided is a medicament for treating a LPA-dependent orLPA-mediated disease or condition in a mammal comprising atherapeutically effective amount of a compound of Formula (I), Formula(II) or Formula (III), or a pharmaceutically acceptable salt thereof.

In some cases disclosed herein is the use of a compound of Formula (I),Formula (II) or Formula (III), or a pharmaceutically acceptable saltthereof, in the manufacture of a medicament for the treatment of aLPA-dependent or LPA-mediated disease or condition.

In some cases disclosed herein is the use of a compound of Formula (I),Formula (II) or Formula (III), or a pharmaceutically acceptable saltthereof in the treatment or prevention of a LPA-dependent orLPA-mediated disease or condition.

In one aspect, is a method for treating or preventing a LPA-dependent orLPA-mediated disease or condition in a mammal comprising administering atherapeutically effective amount of a compound of Formula (I), Formula(II) or Formula (III), or a pharmaceutically acceptable salt thereof.

In one aspect, LPA-dependent or LPA-mediated diseases or conditionsinclude, but are not limited to, fibrosis of organs or tissues,scarring, liver diseases, dermatological conditions, cancer,cardiovascular disease, respiratory diseases or conditions, inflammatorydisease, gastrointestinal tract disease, renal disease, urinarytract-associated disease, inflammatory disease of lower urinary tract,dysuria, frequent urination, pancreas disease, arterial obstruction,cerebral infarction, cerebral hemorrhage, pain, peripheral neuropathy,and fibromyalgia.

In one aspect, the LPA-dependent or LPA-mediated disease or condition isa respiratory disease or condition. In some embodiments, the respiratorydisease or condition is asthma, chronic obstructive pulmonary disease(COPD), pulmonary fibrosis, pulmonary arterial hypertension or acuterespiratory distress syndrome.

In some embodiments, the LPA-dependent or LPA-mediated disease orcondition is selected from idiopathic pulmonary fibrosis; other diffuseparenchymal lung diseases of different etiologies including iatrogenicdrug-induced fibrosis, occupational and/or environmental inducedfibrosis, granulomatous diseases (sarcoidosis, hypersensitivitypneumonia), collagen vascular disease, alveolar proteinosis, langerhanscell granulomatosis, lymphangioleiomyomatosis, inherited diseases(Hermansky-Pudlak Syndrome, tuberous sclerosis, neurofibromatosis,metabolic storage disorders, familial interstitial lung disease);radiation induced fibrosis; chronic obstructive pulmonary disease(COPD); scleroderma; bleomycin induced pulmonary fibrosis; chronicasthma; silicosis; asbestos induced pulmonary fibrosis; acuterespiratory distress syndrome (ARDS); kidney fibrosis;tubulointerstitium fibrosis; glomerular nephritis; focal segmentalglomerular sclerosis; IgA nephropathy; hypertension; Alport; gutfibrosis; liver fibrosis; cirrhosis; alcohol induced liver fibrosis;toxic/drug induced liver fibrosis; hemochromatosis; nonalcoholicsteatohepatitis (NASH); biliary duct injury; primary biliary cirrhosis;infection induced liver fibrosis; viral induced liver fibrosis; andautoimmune hepatitis; corneal scarring; hypertrophic scarring; Duputrendisease, keloids, cutaneous fibrosis; cutaneous scleroderma; spinal cordinjury/fibrosis; myelofibrosis; vascular restenosis; atherosclerosis;arteriosclerosis; Wegener's granulomatosis; Peyronie's disease, chroniclymphocytic leukemia, tumor metastasis, transplant organ rejection,endometreosis, neonatal respiratory distress syndrome and neuropathicpain.

In one aspect, the LPA-dependent or LPA-mediated disease or condition isdescribed herein.

In one aspect, provided is a method for the treatment or prevention oforgan fibrosis in a mammal comprising administering a therapeuticallyeffective amount of a compound of Formula (I), Formula (II) or Formula(III) or a pharmaceutically acceptable salt thereof to a mammal in needthereof.

In one aspect, the organ fibrosis comprises lung fibrosis, renalfibrosis, or hepatic fibrosis.

In one aspect, provided is a method of improving lung function in amammal comprising administering a therapeutically effective amount of acompound of Formula (I), Formula (II) or Formula (III), or apharmaceutically acceptable salt thereof to the mammal in need thereof.In one aspect, the mammal has been diagnosed as having lung fibrosis.

In one aspect, compounds disclosed herein are used to treat idiopathicpulmonary fibrosis (usual interstitial pneumonia) in a mammal.

In some embodiments, compounds disclosed herein are used to treatdiffuse parenchymal interstitial lung diseases in mammal: iatrogenicdrug induced, occupational/environmental (Farmer lung), granulomatousdiseases (sarcoidosis, hypersensitivity pneumonia), collagen vasculardisease (scleroderma and others), alveolar proteinosis, langerhans cellgranulonmatosis, lymphangioleiomyomatosis, Hermansky-Pudlak Syndrome,Tuberous sclerosis, neurofibromatosis, metabolic storage disorders,familial interstitial lung disease.

In some embodiments, compounds disclosed herein are used to treatpost-transplant fibrosis associated with chronic rejection in a mammal:Bronchiolitis obliterans for lung transplant.

In some embodiments, compounds disclosed herein are used to treatcutaneous fibrosis in a mammal: cutaneous scleroderma, Dupuytrendisease, keloids.

In one aspect, compounds disclosed herein are used to treat hepaticfibrosis with or without cirrhosis in a mammal: toxic/drug induced(hemochromatosis), alcoholic liver disease, viral hepatitis (hepatitis Bvirus, hepatitis C virus, HCV), nonalcoholic liver disease (NASH),metabolic and auto-immune.

In one aspect, compounds disclosed herein are used to treat renalfibrosis in a mammal: tubulointerstitium fibrosis, glomerular sclerosis.

In any of the aforementioned aspects involving the treatment of LPAdependent diseases or conditions are further embodiments comprisingadministering at least one additional agent in addition to theadministartion of a compound having the structure of Formula (I),Formula (II) or Formula (III), or a pharmaceutically acceptable saltthereof. In various embodiments, each agent is administered in anyorder, including simultaneously.

In any of the embodiments disclosed herein, the mammal is a human.

In some embodiments, compounds provided herein are administered to ahuman.

In some embodiments, compounds provided herein are orally administered.

In some embodiments, compounds provided herein are used as antagonistsof at least one LPA receptor. In some embodiments, compounds providedherein are used for inhibiting the activity of at least one LPA receptoror for the treatment of a disease or condition that would benefit frominhibition of the activity of at least one LPA receptor. In one aspect,the LPA receptor is LPA₁.

In other embodiments, compounds provided herein are used for theformulation of a medicament for the inhibition of LPA₁ activity.

Articles of manufacture, which include packaging material, a compound ofFormula (I), Formula (II) or Formula (III), or a pharmaceuticallyacceptable salt thereof, within the packaging material, and a label thatindicates that the compound or composition, or pharmaceuticallyacceptable salt, tautomers, pharmaceutically acceptable N-oxide,pharmaceutically active metabolite, pharmaceutically acceptable prodrug,or pharmaceutically acceptable solvate thereof, is used for inhibitingthe activity of at least one LPA receptor, or for the treatment,prevention or amelioration of one or more symptoms of a disease orcondition that would benefit from inhibition of the activity of at leastone LPA receptor, are provided.

Other objects, features and advantages of the compounds, methods andcompositions described herein will become apparent from the followingdetailed description. It should be understood, however, that thedetailed description and the specific examples, while indicatingspecific embodiments, are given by way of illustration only, sincevarious changes and modifications within the spirit and scope of theinstant disclosure will become apparent to those skilled in the art fromthis detailed description.

DETAILED DESCRIPTION OF THE INVENTION

Lysophospholipids are membrane-derived bioactive lipid mediators.Lysophospholipids include, but are not limited to, lysophosphatidic acid(1-acyl-2-hydroxy-sn-glycero-3-phosphate; LPA), sphingosine 1-phosphate(S1P), lysophosphatidylcholine (LPC), and sphingosylphosphorylcholine(SPC). Lysophospholipids affect fundamental cellular functions thatinclude cellular proliferation, differentiation, survival, migration,adhesion, invasion, and morphogensis. These functions influence manybiological processes that include neurogensis, angiogenesis, woundhealing, immunity, and carcinogenesis.

LPA acts through sets of specific G protein-coupled receptors (GPCRs) inan autocrine and paracrine fashion. LPA binding to its cognate GPCRs(LPA₁, LPA₂, LPA₃, LPA₄, LPA₅, LPA₆) activates intracellular signalingpathways to produce a variety of biological responses.

Lysophospholipids, such as LPA, are quantitatively minor lipid speciescompared to their major phospholipid counterparts (e.g.,phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin). LPAhas a role as a biological effector molecule, and has a diverse range ofphysiological actions such as, but not limited to, effects on bloodpressure, platelet activation, and smooth muscle contraction, and avariety of cellular effects, which include cell growth, cell rounding,neurite retraction, and actin stress fiber formation and cell migration.The effects of LPA are predominantly receptor mediated.

Activation of the LPA receptors (LPA₁, LPA₂, LPA₃, LPA₄, LPA₅, LPA₆)with LPA mediates a range of downstream signaling cascades. Theseinclude, but are not limited to, mitogen-activated protein kinase (MAPK)activation, adenylyl cyclase (AC) inhibition/activation, phospholipase C(PLC) activation/Ca²⁺ mobilization, arachidonic acid release, Akt/PKBactivation, and the activation of small GTPases, Rho, ROCK, Rac, andRas. Other pathways that are affected by LPA receptor activationinclude, but are not limited to, cyclic adenosine monophosphate (cAMP),cell division cycle 42/GTP-binding protein (Cdc42), proto-oncogeneserine/threonine-protein kinase Raf (c-RAF), proto-oncogenetyrosine-protein kinase Src (c-src), extracellular signal-regulatedkinase (ERK), focal adhesion kinase (FAK), guanine nucleotide exchangefactor (GEF), glycogen synthase kinase 3b (GSK3b), c-jun amino-terminalkinase (INK), MEK, myosin light chain II (MLC II), nuclear factor kB(NF-kB), N-methyl-D-aspartate (NMDA) receptor activation,phosphatidylinositol 3-kinase (PI3K), protein kinase A (PKA), proteinkinase C (PKC), ras-related C3 botulinum toxin substrate 1 (RAC1). Theactual pathway and realized end point are dependent on a range ofvariables that include receptor usage, cell type, expression level of areceptor or signaling protein, and LPA concentration. Nearly allmammalian cells, tissues and organs co-express several LPA-receptorsubtypes, which indicates that LPA receptors signal in a cooperativemanner. LPA₁, LPA₂, and LPA₃ share high amino acid sequence similarity.

LPA is produced from activated platelets, activated adipocytes, neuronalcells, and other cell types. Serum LPA is produced by multiple enzymaticpathways that involve monoacylglycerol kinase, phospholipase A₁,secretory phospholipase A₂, and lysophospholipase D (lysoPLD), includingautotaxin. Several enzymes are involved in LPA degradation:lysophospholipase, lipid phosphate phosphatase, and LPA acyl transferasesuch as endophilin. LPA concentrations in human serum are estimated tobe 1-5 μM. Serum LPA is bound to albumin, low-density lipoproteins, orother proteins, which possibly protect LPA from rapid degradation. LPAmolecular species with different acyl chain lengths and saturation arenaturally occurring, including 1-palmitoyl (16:0), 1-palmitoleoyl(16:1), 1-stearoyl (18:0), 1-oleoyl (18:1), 1-linoleoyl (18:2), and1-arachidonyl (20:4) LPA. Quantitatively minor alkyl LPA has biologicalactivities similar to acyl LPA, and different LPA species activate LPAreceptor subtypes with varied efficacies.

LPA Receptors

LPA₁ (previously called VZG-1/EDG-2/mrec1.3) couples with three types ofG proteins, G_(i/o), G_(q), and G_(12/13). Through activation of these Gproteins, LPA induces a range of cellular responses through LPA₁including but not limited to: cell proliferation, serum-response element(SRE) activation, mitogen-activated protein kinase (MAPK) activation,adenylyl cyclase (AC) inhibition, phospholipase C (PLC) activation, Ca²mobilization, Akt activation, and Rho activation (Fukushima N, et al.,Annu. Rev Pharmacol. Toxicol. 41:507-34, 2001; Contos J J A, et al.,Mol. Pharmacol. 58:1188-96, 2000).

Wide expression of LPA₁ is observed in adult mice, with clear presencein testis, brain, heart, lung, small intestine, stomach, spleen, thymus,and skeletal muscle. Similarly, human tissues also express LPA₁; it ispresent in brain, heart, lung, placenta, colon, small intestine,prostate, testis, ovary, pancreas, spleen, kidney, skeletal muscle, andthymus.

LPA₂ (EDG-4) also couples with three types of G proteins, G_(i/o),G_(q), and G_(12/13), to mediate LPA-induced cellular signaling.Expression of LPA₂ is observed in the testis, kidney, lung, thymus,spleen, and stomach of adult mice and in the human testis, pancreas,prostate, thymus, spleen, and peripheral blood leukocytes. Expression ofLPA₂ is upregulated in various cancer cell lines, and several human LPA₂transcriptional variants with mutations in the 3′-untranslated regionhave been observed. Targeted deletion of LPA₂ in mice has not shown anyobvious phenotypic abnormalities, but has demonstrated a significantloss of normal LPA signaling (e.g., PLC activation, Ca²⁺ mobilization,and stress fiber formation) in primary cultures of mouse embryonicfibroblasts (MEFs). Creation of lpa1(−/−) lpa2(−/−) double-null mice hasrevealed that many LPA-induced responses, which include cellproliferation, AC inhibition, PLC activation, Ca²⁺ mobilization, JNK andAkt activation, and stress fiber formation, are absent or severelyreduced in double-null MEFs. All these responses, except for ACinhibition (AC inhibition is nearly abolished in LPA₁ (−/−) MEFs), areonly partially affected in either LPA₁ (−/−) or LPA₂ (−/−) MEFs. LPA₂contributes to normal LPA-mediated signaling responses in at least somecell types (Choi et al, Biochemica et Biophysica Acta 2008, 1781, p531-539).

LPA₃ (EDG-7) is distinct from LPA₁ and LPA₂ in its ability to couplewith G_(i/o) and G_(q) but not G_(12/13) and is much less responsive toLPA species with saturated acyl chains. LPA₃ can mediate pleiotropicLPA-induced signaling that includes PLC activation, Ca²⁺ mobilization,AC inhibition/activation, and MAPK activation. Overexpression of LPA₃ inneuroblastoma cells leads to neurite elongation, whereas that of LPA₁ orLPA₂ results in neurite retraction and cell rounding when stimulatedwith LPA. Expression of LPA₃ is observed in adult mouse testis, kidney,lung, small intestine, heart, thymus, and brain. In humans, it is foundin the heart, pancreas, prostate, testis, lung, ovary, and brain(frontal cortex, hippocampus, and amygdala).

LPA₄ (p2y₉/GPR23) is of divergent sequence compared to LPA₁, LPA₂, andLPA₃ with closer similarity to the platelet-activating factor (PAF)receptor. LPA₄ mediates LPA induced Ca²⁺ mobilization and cAMPaccumulation, and functional coupling to the G protein Gs for ACactivation, as well as coupling to other G proteins. The LPA₄ gene isexpressed in the ovary, pancreas, thymus, kidney and skeletal muscle.

LPA₅ (GPR92) is a member of the purinocluster of GPCRs and isstructurally most closely related to LPA₄. LPA₅ is expressed in humanheart, placenta, spleen, brain, lung and gut. LPA₅ also shows very highexpression in the CD8+lymphocyte compartment of the gastrointestinaltract.

LPA₆ (p2y5) is a member of the purinocluster of GPCRs and isstructurally most closely related to LPA₄. LPA₆ is an LPA receptorcoupled to the G12/13-Rho signaling pathways and is expressed in theinner root sheaths of human hair follicles.

Illustrative Biological Activity

Wound Healing

Normal wound healing occurs by a highly coordinated sequence of eventsin which cellular, soluble factors and matrix components act in concertto repair the injury. The healing response can be described as takingplace in four broad, overlapping phases—hemostasis, inflammation,proliferation, and remodeling. Many growth factors and cytokines arereleased into a wound site to initiate and perpetuate wound healingprocesses.

When wounded, damaged blood vessels activate platelets. The activatedplatelets play pivotal roles in subsequent repair processes by releasingbioactive mediators to induce cell proliferation, cell migration, bloodcoagulation, and angiogenesis. LPA is one such mediator that is releasedfrom activated platelets; this induces platelet aggregation along withmitogenic/migration effects on the surrounding cells, such asendothelial cells, smooth muscle cells, fibroblasts, and keratinocytes.

Topical application of LPA to cutaneous wounds in mice promotes repairprocesses (wound closure and increased neoepithelial thickness) byincreasing cell proliferation/migration without affecting secondaryinflammation.

Activation of dermal fibroblasts by growth factors and cytokines leadsto their subsequent migration from the edges of the wound into theprovisional matrix formed by the fibrin clot whereupon the fibroblastsproliferate and start to restore the dermis by secreting and organizingthe characteristic dermal extracellular matrix (ECM). The increasingnumber of fibroblasts within the wound and continuous precipitation ofECM enhances matrix rigidity by applying small tractional forces to thenewly formed granulation tissue. The increase in mechanical stress, inconjunction with transforming growth factor β (TGFβ), induces a-smoothmuscle actin (α-SMA) expression and the subsequent transformation offibroblasts into myofibroblasts. Myofibroblasts facilitate granulationtissue remodeling via myofibroblast contraction and through theproduction of ECM components.

LPA regulates many important functions of fibroblasts in wound healing,including proliferation, migration, differentiation and contraction.Fibroblast proliferation is required in wound healing in order to fillan open wound. In contrast, fibrosis is characterized by intenseproliferation and accumulation of myofibroblasts that activelysynthesize ECM and proinflammatory cytokines. LPA can either increase orsuppress the proliferation of cell types important in wound healing,such as epithelial and endothelial cells (EC), macrophages,keratinocytes, and fibroblasts. A role for LPA₁ in LPA-inducedproliferation was provided by the observation that LPA-stimulatedproliferation of fibroblasts isolated from LPA₁ receptor null mice wasattenuated (Mills et al, Nat Rev. Cancer 2003; 3: 582-591). LPA inducescytoskeletal changes that are integral to fibroblast adhesion,migration, differentiation and contraction.

Fibrosis

Tissue injury initiates a complex series of host wound-healingresponses; if successful, these responses restore normal tissuestructure and function. If not, these responses can lead to tissuefibrosis and loss of function.

For the majority of organs and tissues the development of fibrosisinvolves a multitude of events and factors. Molecules involved in thedevelopment of fibrosis include proteins or peptides (profibroticcytokines, chemokines, metalloproteinases etc.) and phospholipids.Phospholipids involved in the development of fibrosis include plateletactivating factor (PAF), phosphatidyl choline, sphingosine-1 phosphate(S1P) and lysophosphatidic acid (LPA).

A number of muscular dystrophies are characterized by a progressiveweakness and wasting of musculature, and by extensive fibrosis. It hasbeen shown that LPA treatment of cultured myoblasts induced significantexpression of connective tissue growth factor (CTGF). CTGF subsequentlyinduces collagen, fibronectin and integrin expression and inducesdedifferentiation of these myoblasts. Treatment of a variety of celltypes with LPA induces reproducible and high level induction of CTGF (J.P. Pradere, et al., LPA₁ receptor activation promotes renal interstitialfibrosis, J. Am. Soc. Nephrol. 18 (2007) 3110-3118; N. Wiedmaier, etal., Int J Med Microbiol; 298(3-4):231-43, 2008). CTGF is a profibroticcytokine, signaling down-stream and in parallel with TGFβ.

CTGF expression by gingival epithelial cells, which are involved in thedevelopment of gingival fibromatosis, was found to be exacerbated by LPAtreatment (A. Kantarci, et al., J. Pathol. 210 (2006) 59-66).

LPA is associated with the progression of liver fibrosis. In vitro, LPAinduces stellate cell and hepatocyte proliferation. These activatedcells are the main cell type responsible for the accumulation of ECM inthe liver. Furthermore, LPA plasma levels rise during CCl₄-induced liverfibrosis in rodents, or in hepatitis C virus-induced liver fibrosis inhuman (N. Watanabe, et al., Plasma lysophosphatidic acid level and serumautotaxin activity are increased in liver injury in rats in relation toits severity, Life Sci. 81 (2007) 1009-1015; N. Watanabe, et al., J.Clin. Gastroenterol. 41 (2007) 616-623).

An increase of phospholipid concentrations in the bronchoalveolar lavagefluid in rabbits and rodents injected with bleomycin has been reported(K. Kuroda, et al., Phospholipid concentration in lung lavage fluid asbiomarker for pulmonary fibrosis, Inhal. Toxicol. 18 (2006) 389-393; K.Yasuda, et al., Lung 172 (1994) 91-102).

LPA is associated with heart disease and mycocardial remodeling. SerumLPA levels are increased after myocardial infarction in patients and LPAstimulates rat cardiac fibroblast proliferation and collagen production(Chen et al. FEBS Lett. 2006 Aug. 21; 580(19):4737-45).

Pulmonary Fibrosis

In the lung, aberrant wound healing responses to injury contribute tothe pathogenesis of fibrotic lung diseases. Fibrotic lung diseases, suchas idiopathic pulmonary fibrosis (IPF), are associated with highmorbidity and mortality.

LPA is an important mediator of fibroblast recruitment in pulmonaryfibrosis. LPA and LPA₁ play key pathogenic roles in pulmonary fibrosis.Fibroblast chemoattractant activity plays an important role in the lungsin patients with pulmonary fibrosis. Profibrotic effects ofLPA₁-receptor stimulation is explained by LPA₁-receptor-mediatedvascular leakage and increased fibroblast recruitment, both profibroticevents. The LPA-LPA₁ pathway has a role in mediating fibroblastmigration and vascular leakage in IPF. The end result is the aberranthealing process that characterises this fibrotic condition.

The LPA₁ receptor is the LPA receptor most highly expressed onfibroblasts obtained from patients with IPF. Furthermore, BAL obtainedfrom IPF patients induced chemotaxis of human foetal lung fibroblaststhat was blocked by the dual LPA₁-LPA₃ receptor antagonist Ki16425. Inan experimental bleomycin-induced lung injury mouse model, it was shownthat LPA levels were high in bronchoalveolar lavage samples comparedwith unexposed controls. LPA₁ knockout mice are protected from fibrosisafter bleomycin challenge with reduced fibroblast accumulation andvascular leakage. In human subjects with IPF, high LPA levels wereobserved in bronchoalveolar lavage samples compared with healthycontrols. Increased fibroblast chemotactic activity in these samples wasinhibited by the Ki16425 indicating that fibroblast migration ismediated by the LPA-LPA receptor(s) pathway (Tager et al. NatureMedicine, vol. 14, no. 1, 45-54, 2008).

The LPA-LPA₁ pathway is crucial in fibroblast recruitment and vascularleakage in pulmonary fibrosis.

Activation of latent TGF-β by the avβ6 integrin plays a critical role inthe development of lung injury and fibrosis (Munger et al. Cell, vol.96, 319-328, 1999). LPA induces avβ6-mediated TGF-β activation on humanlung epithelial cells (Xu et al. Am. J. Pathology, vol. 174, no. 2,1264-1279, 2009). The LPA-induced avβ6-mediated TGF-β activation ismediated by the LPA2 receptor. Expression of the LPA2 receptor isincreased in epithelial cells and mesenchymal cells in areas of lungfibrosis from IPF patients compared to normal human lung tissue. TheLPA-LPA2 pathway contributes to the activation of the TGF-β pathway inpulmonary fibrosis. In some embodiments, compounds that inhibit LPA2show efficacy in the treatment of lung fibrosis. In some embodiments,compounds that inhibit both LPA1 and LPA2 show improved efficacy in thetreatment of lung fibrosis compared to compounds which inhibit only LPA1or LPA2.

Renal Fibrosis

LPA and LPA₁ are involved in the etiology of kidney fibrosis. LPA haseffects on both proliferation and contraction of glomerular mesangialcells and thus has been implicated in proliferative glomerulonephritis(C. N. Inoue, et al., Clin. Sci. (Colch.) 96, 431-436, 1999). In ananimal model of renal fibrosis, unilateral ureteral obstruction (UUO),it was found that renal LPA receptors are expressed under basalconditions with an expression order of LPA₂>LPA₃=LPA₁>>LPA₄. This modelmimics in an accelerated manner the development of renal fibrosisincluding renal inflammation, fibroblast activation and accumulation ofextracellular matrix in the tubulointerstitium. UUO significantlyinduced LPA₁-receptor expression. This was paralleled by renal LPAproduction (3.3 fold increase) in conditioned media from kidneyexplants. Contra-lateral kidneys exhibited no significant changes in LPArelease and LPA-receptors expression. This shows that a prerequisite foran action of LPA in fibrosis is met: production of a ligand (LPA) andinduction of one of its receptors (the LPA₁ receptor) (J. P. Pradere etal., Biochimica et Biophysica Acta (2008),doi:10.1016/j.bbalip.2008.04.001).

In mice invalidated for the LPA₁ receptor (LPA₁ (−/−), the developmentof renal fibrosis was significantly attenuated. UUO mice treated withthe LPA receptor antagonist Ki16425 closely resembled the LPA₁ (−/−)mice.

LPA can participate in intraperitonial accumulation ofmonocyte/macrophages and that LPA can induce expression of theprofibrotic cytokine CTGF in primary cultures of human fibroblasts (J.S. Koh, et al., J. Clin. Invest. 102 (1998) 716-727).

LPA treatment of a mouse epithelial renal cell line, MCT, induced arapid increase in the expression of the profibrotic cytokine CTGF. CTGFplays a crucial role in UUO-induced tubulointerstitial fibrosis (TIF),and is involved in the profibrotic activity of TGFβ. This induction wasalmost completely suppressed by co-treatment with the LPA-receptorantagonist Ki16425. In one aspect, the profibrotic activity of LPA inkidney results from a direct action of LPA on kidney cells involvinginduction of CTGF.

Hepatic Fibrosis

LPA is implicated in liver disease and fibrosis. Plasma LPA levels andserum autotoxin (enzyme responsible for LPA production) are elevated inhepatitis patients and animal models of liver injury in correlation withincreased fibrosis. LPA also regulates liver cell function. LPA₁ andLPA₂ receptors are expressed by mouse hepatic stellate cells and LPAstimulates migration of hepatic myofibroblasts.

Ocular Fibrosis

LPA is in involved in wound healing in the eye. LPA₁ and LPA₃ receptorsare detectable in the normal rabbit corneal epithelial cells,keratocytes and endothelial cells and LPA₁ and LPA₃ expression areincreased in corneal epithelial cells following injury.

LPA and its homologues are present in the aqueous humor and the lacrimalgland fluid of the rabbit eye and these levels are increased in a rabbitcorneal injury model.

LPA induces actin stress fiber formation in rabbit corneal endothelialand epithelial cells and promotes contraction corneal fibroblasts. LPAalso stimulates proliferation of human retinal pigmented epithelialcells

Cardiac Fibrosis

LPA is implicated in myocardial infarction and cardiac fibrosis. SerumLPA levels are increased in patients following mycocardial infarction(MI) and LPA stimulates proliferation and collagen production (fibrosis)by rat cardiac fibroblasts. Both LPA1 and LPA3 receptors are highlyexpressed in human heart tissue.

Treatment of Fibrosis

In one aspect, compounds of Formula (I), Formula (II) or Formula (III)are used to treat or prevent fibrosis in a mammal. In one aspect,compounds of Formula (I), Formula (II) or Formula (III) are used totreat fibrosis of an organ or tissue in a mammal. In one aspect is amethod for preventing a fibrosis condition in a mammal, the methodcomprising administering to the mammal at risk of developing one or morefibrosis conditions a therapeutically effective amount of a compound ofFormula (I), Formula (II) or Formula (III). In one aspect, the mammalhas been exposed to one or more environmental conditions that are knownto increase the risk of fibrosis of an organ or tissue. In one aspect,the mammal has been exposed to one or more environmental conditions thatare known to increase the risk of lung, liver or kidney fibrosis. In oneaspect, the mammal has a genetic predisposition of developing fibrosisof an organ or tissue. In one aspect, compounds of Formula (I), Formula(II) and Formula (III) are administered to a mammal to prevent orminimize scarring following injury. In one aspect, injury includessurgery.

The terms “fibrosis” or “fibrosing disorder,” as used herein, refers toconditions that are associated with the abnormal accumulation of cellsand/or fibronectin and/or collagen and/or increased fibroblastrecruitment and include but are not limited to fibrosis of individualorgans or tissues such as the heart, kidney, liver, joints, lung,pleural tissue, peritoneal tissue, skin, cornea, retina, musculoskeletaland digestive tract.

Exemplary diseases, disorders, or conditions that involve fibrosisinclude, but are not limited to: Lung diseases associated with fibrosis,e.g., idiopathic pulmonary fibrosis, pulmonary fibrosis secondary tosystemic inflammatory disease such as rheumatoid arthritis, scleroderma,lupus, cryptogenic fibrosing alveolitis, radiation induced fibrosis,chronic obstructive pulmonary disease (COPD), scleroderma, chronicasthma, silicosis, asbestos induced pulmonary or pleural fibrosis, acutelung injury and acute respiratory distress (including bacterialpneumonia induced, trauma induced, viral pneumonia induced, ventilatorinduced, non-pulmonary sepsis induced, and aspiration induced); Chronicnephropathies associated with injury/fibrosis (kidney fibrosis), e.g.,glomerulonephritis secondary to systemic inflammatory diseases such aslupus and scleroderma, diabetes, glomerular nephritis, focal segmentalglomerular sclerosis, IgA nephropathy, hypertension, allograft andAlport; Gut fibrosis, e.g., scleroderma, and radiation induced gutfibrosis; Liver fibrosis, e.g., cirrhosis, alcohol induced liverfibrosis, nonalcoholic steatohepatitis (NASH), biliary duct injury,primary biliary cirrhosis, infection or viral induced liver fibrosis(e.g., chronic HCV infection), and autoimmune hepatitis; Head and neckfibrosis, e.g., radiation induced; Corneal scarring, e.g., LASIK(laser-assisted in situ keratomileusis), corneal transplant, andtrabeculectomy; Hypertrophic scarring and keloids, e.g., burn induced orsurgical; and Other fibrotic diseases, e.g., sarcoidosis, scleroderma,spinal cord injury/fibrosis, myelofibrosis, vascular restenosis,atherosclerosis, arteriosclerosis, Wegener's granulomatosis, mixedconnective tissue disease, and Peyronie's disease.

In one aspect, a mammal suffering from one of the following non-limitingexemplary diseases, disorders, or conditions will benefit from therapywith a compound of Formula (I), Formula (II) or Formula (III):atherosclerosis, thrombosis, heart disease, vasculitis, formation ofscar tissue, restenosis, phlobitis, COPD (chronic obstructive pulmonarydisease), pulmonary hypertension, pulmonary fibrosis, pulmonaryinflammation, bowel adhesions, bladder fibrosis and cystitis, fibrosisof the nasal passages, sinusitis, inflammation mediated by neutrophils,and fibrosis mediated by fibroblasts.

In one aspect, compounds of Formula (I), Formula (II) and Formula (III),are administered to a mammal with fibrosis of an organ or tissue or witha predisposition of developing fibrosis of an organ or tissue with oneor more other agents that are used to treat fibrosis. In one aspect, theone or more agents include corticosteroids. In one aspect, the one ormore agents include immunosuppresants. In one aspect, the one or moreagents include B-cell antagonists. In one aspect, the one or more agentsinclude uteroglobin.

In one aspect, compounds of Formula (I), Formula (II) and Formula (III),are used to treat a dermatological disorders in a mammal. The term“dermatological disorder,” as used herein refers to a skin disorder.Such dermatological disorders include, but are not limited to,proliferative or inflammatory disorders of the skin such as, atopicdermatitis, bullous disorders, collagenoses, psoriasis, psoriaticlesions, dermatitis, contact dermatitis, eczema, urticaria, rosacea,wound healing, scarring, hypertrophic scarring, keloids, KawasakiDisease, rosacea, Sjogren-Larsso Syndrome, urticaria.

Pain

Since LPA is released following tissue injury, LPA₁ plays an importantrole in the initiation of neuropathic pain. LPA₁, unlike LPA₂ or LPA₃,is expressed in both dorsal root ganglion (DRG) and dorsal root neurons.Using the antisense oligodeoxynucleotide (AS-ODN) for LPA₁ and LPA₁-nullmice, it was found that LPA-induced mechanical allodynia andhyperalgesia is mediated in an LPA₁-dependent manner. LPA₁ anddownstream Rho-ROCK activation play a role in the initiation ofneuropathic pain signaling. Pretreatment with Clostridium botulinum C3exoenzyme (BoTXC3, Rho inhibitor) or Y-27632 (ROCK inhibitor) completelyabolished the allodynia and hyperalgesia in nerve-injured mice. LPA alsoinduced demyelination of the dorsal root, which was prevented by BoTXC3.The dorsal root demyelination by injury was not observed in LPA₁-nullmice or AS-ODN injected wild-type mice. LPA signaling appears to induceimportant neuropathic pain markers such as protein kinase Cγ (PKCγ) anda voltage-gated calcium channel α2δ1 subunit (Caα2δ1) in an LPA₁ andRho-dependent manner (M. Inoue, et al., Initiation of neuropathic painrequires lysophosphatidic acid receptor signaling, Nat. Med. 10 (2004)712-718).

In one aspect, compounds of Formula (I), Formula (II) or Formula (III),are used in the treatment of pain in a mammal. In one aspect, the painis acute pain or chronic pain. In another aspect, the pain isneuropathic pain. In another aspect, the pain is cancer pain.

In one aspect, compounds of Formula (I), Formula (II) and Formula (III),are used in the treatment of fibromylagia. In one aspect, fibromyalgiastems from the formation of fibrous scar tissue in contractile(voluntary) muscles. Fibrosis binds the tissue and inhibits blood flow,resulting in pain.

Cancer

Lysophospholipid receptor signaling plays a role in the etiology ofcancer. Lysophosphatidic acid (LPA) and its G protein-coupled receptors(GPCRs) LPA₁, LPA₂, and/or LPA₃ play a role in the development ofseveral types of cancers. The initiation, progression and metastasis ofcancer involve several concurrent and sequential processes includingcell proliferation and growth, survival and anti-apoptosis, migration ofcells, penetration of foreign cells into defined cellular layers and/ororgans, and promotion of angiogenesis. The control of each of theseprocesses by LPA signaling in physiological and pathophysiologicalconditions underscores the potential therapeutic usefulness ofmodulating LPA signaling pathways for the treatment of cancer,especially at the level of the LPA receptors or ATX/lysoPLD. Autotaxin(ATX) is a prometastatic enzyme initially isolated from the conditionedmedium of human melanoma cells that stimulates a myriad of biologicalactivities, including angiogenesis and the promotion of cell growth,migration, survival, and differentiation through the production of LPA(Mol Cancer Ther 2008; 7(10):3352-62).

LPA signals through its own GPCRs leading to activation of multipledownstream effector pathways. Such downstream effector pathways play arole in cancer. LPA and its GPCRs are linked to cancer through majoroncogenic signaling pathways.

LPA contributes to tumorigenesis by increasing motility and invasivenessof cells. LPA has been implicated in the initiation or progression ofovarian cancer. LPA is present at significant concentrations (2-80 μM)in the ascitic fluid of ovarian cancer patients. Ovarian cancer cellsconstitutively produce increased amounts of LPA as compared to normalovarian surface epithelial cells, the precursor of ovarian epithelialcancer. Elevated LPA levels are also detected in plasma from patientswith early-stage ovarian cancers compared with controls. LPA receptors(LPA2 and LPA3) are also overexpressed in ovarian cancer cells ascompared to normal ovarian surface epithelial cells. LPA stimulatesCox-2 expression through transcriptional activation andpost-transcriptional enhancement of Cox-2 mRNA in ovarian cancer cells.Prostaglandins produced by Cox-2 have been implicated in a number ofhuman cancers and pharmacological inhibition of Cox-2 activity reducescolon cancer development and decreases the size and number of adenomasin patients with familial adenomatous polyposis. LPA has also beenimplicated in the initiation or progression of prostate cancer, breastcancer, melanoma, head and neck cancer, bowel cancer (colorectalcancer), thyroid cancer, glioblastoma, and other cancers (Gardell et al,Trends in Molecular Medicine, vol. 12, no. 2, p 65-75, 2006; Ishii etal, Annu. Rev. Biochem, 73, 321-354, 2004; Mills et al., Nat. Rev.Cancer, 3, 582-591, 2003; Murph et al., Biochimica et Biophysica Acta,1781, 547-557, 2008; Kishi et al., J. Biol. Chem., 281, 17492-17500,2006).

The cellular responses to LPA are mediated through the lysophosphatidicacid receptors. For example, LPA receptors mediate both migration of andinvasion by pancreatic cancer cell lines: an antagonist of LPA₁ and LPA₃(Ki16425) and LPA₁-specific siRNA effectively blocked in vitro migrationin response to LPA and peritoneal fluid (ascites) from pancreatic cancerpatients; in addition, Ki16425 blocked the LPA-induced andascites-induced invasion activity of a highly peritoneal metastaticpancreatic cancer cell line (Yamada et al, J. Biol. Chem., 279,6595-6605, 2004).

Colorectal carcinoma cell lines show significant expression of LPA₁ mRNAand respond to LPA by cell migration and production of angiogenicfactors. Overexpression of LPA receptors has a role in the pathogenesisof thyroid cancer. LPA₃ was originally cloned from prostate cancercells, concordant with the ability of LPA to induce autocrineproliferation of prostate cancer cells.

LPA has stimulatory roles in cancer progression in many types of cancer.LPA is produced from and induces proliferation of prostate cancer celllines. LPA induces human colon carcinoma DLD1 cell proliferation,migration, adhesion, and secretion of angiogenic factors through LPA₁signalling. In other human colon carcinoma cells lines (HT29 and WiDR),LPA enhances cell proliferation and secretion of angiogenic factors. Inother colon cancer cell lines, LPA₂ and LPA₃ receptor activation resultsin proliferation of the cells. LPA₁ is implicated in bone metastasis andthe LPA₁/LPA₃ dual antagonist Ki16425 has been shown to inhibitmetastasis to bone in vivo (Boucharaba et al., Proc. Natl. Acad. SciUSA, 103, 9643-9648, 2006). The genetic or pharmacological manipulationof LPA metabolism, specific blockade of receptor signaling, and/orinhibition of downstream signal transduction pathways, representapproaches for cancer therapies.

In one aspect, a compound of Formula (I), Formula (II) or Formula (III),is used in the treatment of cancer. In one aspect, compounds of Formula(I), Formula (II) or Formula (III), are used in the treatment ofmalignant and benign proliferative disease. In one aspect, compounds ofFormula (I), Formula (II) and Formula (III) are used to prevent orreduce proliferation of tumor cells, invasion and metastasis ofcarcinomas, pleural mesothelioma (Yamada, Cancer Sci., 2008, 99(8),1603-1610) or peritoneal mesothelioma, cancer pain, bone metastases(Boucharaba et al, J. Clin. Invest., 2004, 114(12), 1714-1725;Boucharaba et al, Proc. Natl. acad. Sci., 2006, 103(25) 9643-9648). Inone aspect is a method of treating cancer in a mammal, the methodcomprising administering to the mammal a compound of Formula (I),Formula (II) or Formula (III) and a second therapeutic agent, whereinthe second therapeutic agent is an anti-cancer agent.

The term “cancer,” as used herein refers to an abnormal growth of cellswhich tend to proliferate in an uncontrolled way and, in some cases, tometastasize (spread). The types of cancer include, but is not limitedto, solid tumors (such as those of the bladder, bowel, brain, breast,endometrium, heart, kidney, lung, lymphatic tissue (lymphoma), ovary,pancreas or other endocrine organ (thyroid), prostate, skin (melanoma orbasal cell cancer) or hematological tumors (such as the leukemias) atany stage of the disease with or without metastases.

The increased concentrations of LPA and vesicles in ascites from ovariancancer patients and breast cancer effussions indicate that it could bean early diagnostic marker, a prognostic indicator or an indicator ofresponse to therapy (Mills et al, Nat. Rev. Cancer., 3, 582-591, 2003;Sutphen et al., Cancer Epidemiol. Biomarkers Prey. 13, 1185-1191, 2004).LPA concentrations are consistently higher in ascites samples than inmatched plasma samples.

Respiratory and Allergic Disorders

In one aspect, LPA is a contributor to the pathogenesis of respiratorydiseases. In one aspect the respiratory disease is asthma.Proinflammatory effects of LPA include degranulation of mast cells,contraction of smooth-muscle cells and release of cytokines fromdendritic cells. Airway smooth muscle cells, epithelial cells and lungfibroblasts all show responses to LPA. LPA induces the secretion of IL-8from human bronchial epithelial cells. IL-8 is found in increasedconcentrations in BAL fluids from patients with asthma, chronicobstructive lung disease, pulmonary sarcoidosis and acute respiratorydistress syndrome and Il-8 has been shown to exacerbate airwayinflammation and airway remodeling of asthmatics. LPA1, LPA2 and LPA3receptors have all been shown to contribute to the LPA-induced IL-8production. Studies cloning multiple GPCRs that are activated by LPAallowed the demonstration of the presence of mRNA for the LPA₁, LPA₂ andLPA₃ in the lung (J. J. A. Contos, et al., Mol. Pharmacol. 58,1188-1196, 2000).

The release of LPA from platelets activated at a site of injury and itsability to promote fibroblast proliferation and contraction are featuresof LPA as a mediator of wound repair. In the context of airway disease,asthma is an inflammatory disease where inappropriate airway “repair”processes lead to structural “remodeling” of the airway. In asthma, thecells of the airway are subject to ongoing injury due to a variety ofinsults, including allergens, pollutants, other inhaled environmentalagents, bacteria and viruses, leading to the chronic inflammation thatcharacterizes asthma.

In one aspect, in the asthmatic individual, the release of normal repairmediators, including LPA, is exaggerated or the actions of the repairmediators are inappropriately prolonged leading to inappropriate airwayremodeling. Major structural features of the remodeled airway observedin asthma include a thickened lamina reticularis (the basementmembrane-like structure just beneath the airway epithelial cells),increased numbers and activation of myofibroblasts, thickening of thesmooth muscle layer, increased numbers of mucus glands and mucussecretions, and alterations in the connective tissue and capillary bedthroughout the airway wall. In one aspect, LPA contributes to thesestructural changes in the airway. In one aspect, LPA is involved inacute airway hyperresponsiveness in asthma. The lumen of the remodeledasthmatic airway is narrower due to the thickening of the airway wall,thus decreasing airflow. In one aspect, LPA contributes to the long-termstructural remodeling and the acute hyperresponsiveness of the asthmaticairway. In one aspect, LPA contributes to the hyper-responsiveness thatis a primary feature of acute exacerbations of asthma.

In addition to the cellular responses mediated by LPA, several of theLPA signaling pathway components leading to these responses are relevantto asthma. EGF receptor upregulation is induced by LPA and is also seenin asthmatic airways (M. Amishima, et al., Am. J. Respir. Crit. CareMed. 157, 1907-1912, 1998). Chronic inflammation is a contributor toasthma, and several of the transcription factors that are activated byLPA are known to be involved in inflammation (Ediger et al., Eur RespirJ 21:759-769, 2003).

In one aspect, the fibroblast proliferation and contraction andextracellular matrix secretion stimulated by LPA contributes to thefibroproliferative features of other airway diseases, such as theperibronchiolar fibrosis present in chronic bronchitis, emphysema, andinterstitial lung disease. Emphysema is also associated with a mildfibrosis of the alveolar wall, a feature which is believed to representan attempt to repair alveolar damage. In another aspect, LPA plays arole in the fibrotic interstitial lung diseases and obliterativebronchiolitis, where both collagen and myofibroblasts are increased. Inanother aspect, LPA is involved in several of the various syndromes thatconstitute chronic obstructive pulmonary disease.

Administration of LPA in vivo induces airway hyper-responsiveness,itch-scratch responses, infiltration and activation of eosinophils andneutrophils, vascular remodeling, and nociceptive flexor responses. LPAalso induces histamine release from mouse and rat mast cells. In anacute allergic reaction, histamine induces various responses, such ascontraction of smooth muscle, plasma exudation, and mucus production.Plasma exudation is important in the airway, because the leakage andsubsequent airway-wall edema contribute to the development of airwayhyperresponsiveness. Plasma exudation progresses to conjunctivalswelling in ocular allergic disorder and nasal blockage in allergicrhinitis (Hashimoto et al., J Pharmacol Sci 100, 82-87, 2006). In oneaspect, plasma exudation induced by LPA is mediated by histamine releasefrom mast cells via one or more LPA receptors. In one aspect, the LPAreceptor(s) include LPA₁ and/or LPA₃. In one aspect, compounds ofFormula (I), Formula (II) and Formula (III) are used in the treatment ofvarious allergic disorders in a mammal. In one aspect, compounds ofFormula (I), Formula (II) and Formula (III) are used in the treatment ofrespiratory diseases, disorders or conditions in a mammal. In oneaspect, compounds of Formula (I), Formula (II) or Formula (III) are usedin the treatment of asthma in a mammal. In one aspect, compounds ofFormula (I), Formula (II) and Formula (III) are used in the treatment ofchronic asthma in a mammal.

The term “respiratory disease,” as used herein, refers to diseasesaffecting the organs that are involved in breathing, such as the nose,throat, larynx, eustachian tubes, trachea, bronchi, lungs, relatedmuscles (e.g., diaphram and intercostals), and nerves. Respiratorydiseases include, but are not limited to, asthma, adult respiratorydistress syndrome and allergic (extrinsic) asthma, non-allergic(intrinsic) asthma, acute severe asthma, chronic asthma, clinicalasthma, nocturnal asthma, allergen-induced asthma, aspirin-sensitiveasthma, exercise-induced asthma, isocapnic hyperventilation, child-onsetasthma, adult-onset asthma, cough-variant asthma, occupational asthma,steroid-resistant asthma, seasonal asthma, seasonal allergic rhinitis,perennial allergic rhinitis, chronic obstructive pulmonary disease,including chronic bronchitis or emphysema, pulmonary hypertension,interstitial lung fibrosis and/or airway inflammation and cysticfibrosis, and hypoxia.

The term “asthma” as used herein refers to any disorder of the lungscharacterized by variations in pulmonary gas flow associated with airwayconstriction of whatever cause (intrinsic, extrinsic, or both; allergicor non-allergic). The term asthma may be used with one or moreadjectives to indicate cause.

In one aspect, presented herein is the use of compounds of Formula (I),Formula (II) and Formula (III) in the treatment or prevention of chronicobstructive pulmonary disease in a mammal comprising administering tothe mammal at least once an effective amount of at least one compound ofFormula (I), Formula (II) or Formula (III). In addition, chronicobstructive pulmonary disease includes, but is not limited to, chronicbronchitis or emphysema, pulmonary hypertension, interstitial lungfibrosis and/or airway inflammation, and cystic fibrosis.

Nervous System

The nervous system is a major locus for LPA₁ expression; there it isspatially and temporally regulated throughout brain development.Oligodendrocytes, the myelinating cells in the central nervous system(CNS), express LPA₁ in mammals. In addition, Schwann cells, themyelinating cells of the peripheral nervous system, also express LPA₁,which is involved in regulating Schwann cell survival and morphology.These observations identify important functions for receptor-mediatedLPA signaling in neurogenesis, cell survival, and myelination.

Exposure of peripheral nervous system cell lines to LPA produces a rapidrefraction of their processes resulting in cell rounding, which was, inpart, mediated by polymerization of the actin cytoskeleton. In oneaspect, LPA causes neuronal degeneration under pathological conditionswhen the blood-brain barrier is damaged and serum components leak intothe brain (Moolenaar, Curr. Opin. Cell Biol. 7:203-10, 1995).Immortalized CNS neuroblast cell lines from the cerebral cortex alsodisplay retraction responses to LPA exposure through Rho activation andactomyosin interactions. In one aspect, LPA is associated withpost-ischemic neural damage (J. Neurochem. 61, 340, 1993; J. Neurochem.,70:66, 1998).

In one aspect, provided is a compound of Formula (I), Formula (II) orFormula (III) for use in the treatment or prevention of a nervous systemdisorder in a mammal. The term “nervous system disorder,” as usedherein, refers to conditions that alter the structure or function of thebrain, spinal cord or peripheral nervous system, including but notlimited to Alzheimer's Disease, cerebral edema, cerebral ischemia,stroke, multiple sclerosis, neuropathies, Parkinson's Disease, thosefound after blunt or surgical trauma (including post-surgical cognitivedysfunction and spinal cord or brain stem injury), as well as theneurological aspects of disorders such as degenerative disk disease andsciatica.

In one aspect, provided is a compound of Formula (I), Formula (II) orFormula (III) for use in the treatment or prevention of a CNS disorderin a mammal. CNS disorders include, but are not limited to, multiplesclerosis, Parkinson's disease, Alzheimer's disease, stroke, cerebralischemia, retinal ischemia, post-surgical cognitive dysfunction,migraine, peripheral neuropathy/neuropathic pain, spinal cord injury,cerebral edema and head injury.

Cardiovascular Disorders

Cardiovascular phenotypes observed after targeted deletion oflysophospholipid receptors reveal important roles for lysophospholipidsignaling in the development and maturation of blood vessels, formationof atherosclerotic plaques and maintenance of heart rate (Ishii, I. etal. Annu. Rev. Biochem. 73, 321-354, 2004). Angiogenesis, the formationof new capillary networks from pre-existing vasculature, is normallyinvoked in wound healing, tissue growth and myocardial angiogenesisafter ischemic injury. Peptide growth factors (e.g. vascular endothelialgrowth factor (VEGF)) and lysophospholipids control coordinatedproliferation, migration, adhesion, differentiation and assembly ofvascular endothelial cells (VECs) and surrounding vascular smooth-musclecells (VSMCs). In one aspect, dysregulation of the processes mediatingangiogenesis leads to atherosclerosis, hypertension, tumor growth,rheumatoid arthritis and diabetic retinopathy (Osborne, N. and Stainier,D. Y. Annu. Rev. Physiol. 65, 23-43, 2003).

Downstream signaling pathways evoked by lysophospholipid receptorsinclude Rac-dependent lamellipodia formation (e.g. LPA₁) andRho-dependent stress-fiber formation (e.g. LPA₁), which is important incell migration and adhesion. Dysfunction of the vascular endothelium canshift the balance from vasodilatation to vasoconstriction and lead tohypertension and vascular remodeling, which are risk factors foratherosclerosis (Maguire, J. J. et al., Trends Pharmacol. Sci. 26,448-454, 2005).

LPA contributes to both the early phase (barrier dysfunction andmonocyte adhesion of the endothelium) and the late phase (plateletactivation and intra-arterial thrombus formation) of atherosclerosis, inaddition to its overall progression. In the early phase, LPA fromnumerous sources accumulates in lesions and activates its cognate GPCRs(LPA₁ and LPA₃) expressed on platelets (Siess, W. Biochim. Biophys. Acta1582, 204-215, 2002; Rother, E. et al. Circulation 108, 741-747, 2003).This triggers platelet shape change and aggregation, leading tointra-arterial thrombus formation and, potentially, myocardialinfarction and stroke. In support of its atherogenic activity, LPA canalso be a mitogen and motogen to VSMCs and an activator of endothelialcells and macrophages. LPA has been shown to be involved inischemia-reperfussion injury. Blockade of the LPA₃ receptor in a murinemodel of renal ischemia-reperfusion injury reduced the severity ofinjury. This effect was reversed in the presence of the selective LPA₃receptor agonist OMPT (Okusa et al., Am. J. Physiol. Renal Physiol.,285, F565-F574, 2003). In one aspect, mammals with cardiovasculardisease benefit from LPA receptor antagonists that prevent thrombus andneointima plaque formation.

The specific effects of LPA are receptor-mediated.

In one aspect, compounds of Formula (I), Formula (II) and Formula (III)are used to treat or prevent cardiovascular disease in mammal.

The term “cardiovascular disease,” as used herein refers to diseasesaffecting the heart or blood vessels or both, including but not limitedto: arrhythmia (atrial or ventricular or both); atherosclerosis and itssequelae; angina; cardiac rhythm disturbances; myocardial ischemia;myocardial infarction; cardiac or vascular aneurysm; vasculitis, stroke;peripheral obstructive arteriopathy of a limb, an organ, or a tissue;reperfusion injury following ischemia of the brain, heart, kidney orother organ or tissue; endotoxic, surgical, or traumatic shock;hypertension, valvular heart disease, heart failure, abnormal bloodpressure; shock; vasoconstriction (including that associated withmigraines); vascular abnormality, inflammation, insufficiency limited toa single organ or tissue.

In one aspect, provided herein are methods for preventing or treatingvasoconstriction, atherosclerosis and its sequelae myocardial ischemia,myocardial infarction, aortic aneurysm, vasculitis and stroke comprisingadministering at least once to the mammal an effective amount of atleast one compound of Formula (I), Formula (II) or Formula (III) orpharmaceutical composition or medicament which includes a compound ofFormula (I), Formula (II) or Formula (III).

In one aspect, provided herein are methods for reducing cardiacreperfusion injury following myocardial ischemia and/or endotoxic shockcomprising administering at least once to the mammal an effective amountof at least one compound of Formula (I), Formula (II) or Formula (III).

In one aspect, provided herein are methods for reducing the constrictionof blood vessels in a mammal comprising administering at least once tothe mammal an effective amount of at least one compound of Formula (I),Formula (II) or Formula (III).

In one aspect, provided herein are methods for lowering or preventing anincrease in blood pressure of a mammal comprising administering at leastonce to the mammal an effective amount of at least one compound ofFormula (I), Formula (II) or Formula (III).

Inflammation

LPA has been shown to regulate immunological responses by modulatingactivities/functions of immune cells such as T-/B-lymphocytes andmacrophages. In activated T cells, LPA activates IL-2 production/cellproliferation through LPA₁ (Gardell et al, TRENDS in Molecular MedicineVol. 12 No. 2 February 2006). Expression of LPA-induced inflammatoryresponse genes is mediated by LPA₁ and LPA₃ (Biochem Biophys Res Commun.363(4):1001-8, 2007). In addition, LPA modulates the chemotaxis ofinflammatory cells (Biochem Biophys Res Commun., 1993, 15; 193(2), 497).The proliferation and cytokine-secreting activity in response to LPA ofimmune cells (J. Imuunol. 1999, 162, 2049), platelet aggregationactivity in response to LPA, acceleration of migration activity inmonocytes, activation of NF-κB in fibroblast, enhancement offibronectin-binding to the cell surface, and the like are known. Thus,LPA is associated with various inflammatory/immune diseases.

In one aspect, compounds of Formula (I), Formula (II) and Formula (III)are used to treat or prevent inflammation in a mammal. In one aspect,antagonists of LPA₁ and/or LPA₃ find use in the treatment or preventionof inflammatory/immune disorders in a mammal. In one aspect, theantagonist of LPA₁ is a compound of Formula (I), Formula (II) or Formula(III).

Examples of inflammatory/immune disorders include psoriasis, rheumatoidarthritis, vasculitis, inflammatory bowel disease, dermatitis,osteoarthritis, asthma, inflammatory muscle disease, allergic rhinitis,vaginitis, interstitial cystitis, scleroderma, eczema, allogeneic orxenogeneic transplantation (organ, bone marrow, stem cells and othercells and tissues) graft rejection, graft-versus-host disease, lupuserythematosus, inflammatory disease, type I diabetes, pulmonaryfibrosis, dermatomyositis, Sjogren's syndrome, thyroiditis (e.g.,Hashimoto's and autoimmune thyroiditis), myasthenia gravis, autoimmunehemolytic anemia, multiple sclerosis, cystic fibrosis, chronic relapsinghepatitis, primary biliary cirrhosis, allergic conjunctivitis and atopicdermatitis.

Other Diseases, Disorders or Conditions

In accordance with one aspect, are methods for treating, preventing,reversing, halting or slowing the progression of LPA-dependent orLPA-mediated diseases or conditions once it becomes clinically evident,or treating the symptoms associated with or related to LPA-dependent orLPA-mediated diseases or conditions, by administering to the mammal acompound of Formula (I), Formula (II) or Formula (III). In certainembodiments, the subject already has a LPA-dependent or LPA-mediateddisease or condition at the time of administration, or is at risk ofdeveloping a LPA-dependent or LPA-mediated disease or condition.

In certain aspects, the activity of LPA₁ in a mammal is directly orindirectly modulated by the administration of (at least once) atherapeutically effective amount of at least one compound of Formula(I), Formula (II) or Formula (III). Such modulation includes, but is notlimited to, reducing and/or inhibiting the activity of LPA₁. Inadditional aspects, the activity of LPA in a mammal is directly orindirectly modulated, including reducing and/or inhibiting, by theadministration of (at least once) a therapeutically effective amount ofat least one compound of Formula (I), Formula (II) or Formula (III).Such modulation includes, but is not limited to, reducing and/orinhibiting the amount and/or activity of a LPA receptor. In one aspect,the LPA receptor is LPA₁.

In one aspect, LPA has a contracting action on bladder smooth musclecell isolated from bladder, and promotes growth of prostate-derivedepithelial cell (The Journal of Urology, 1999, vol. 162, pp. 1779-1784;The Journal of Urology, 2000, vol. 163, pp. 1027-1032). In anotheraspect, LPA contracts the urinary tract and prostate in vitro andincreases intraurethral pressure in vivo (WO 02/062389).

In certain aspects, are methods for preventing or treating eosinophiland/or basophil and/or dendritic cell and/or neutrophil and/or monocyteand/or T-cell recruitment comprising administering at least once to themammal an effective amount of at least one compound of Formula (I),Formula (II) or Formula (III).

In certain aspects, are methods for the treatment of cystitis,including, e.g.,interstitial cystitis, comprising administering at leastonce to the mammal a therapeutically effective amount of at least onecompound of Formula (I), Formula (II) or Formula (III).

In accordance with one aspect, methods described herein include thediagnosis or determination of whether or not a patient is suffering froma LPA-dependent or LPA-mediated disease or condition by administering tothe subject a therapeutically effective amount of a compound of Formula(I), Formula (II) or Formula (III) and determining whether or not thepatient responds to the treatment.

In one aspect provided herein are compounds of Formula (I), Formula (II)and Formula (III), pharmaceutically acceptable salts, pharmaceuticallyacceptable prodrugs, and pharmaceutically acceptable solvates thereof,which are antagonists of LPA₁, and are used to treat patients sufferingfrom one or more LPA-dependent or LPA-mediated conditions or diseases,including, but not limited to, lung fibrosis, kindney fibrosis, liverfibrosis, scarring, asthma, rhinitis, chronic obstructive pulmonarydisease, pulmonary hypertension, interstitial lung fibrosis, arthritis,allergy, psoriasis, inflammatory bowel disease, adult respiratorydistress syndrome, myocardial infarction, aneurysm, stroke, cancer,pain, proliferative disorders and inflammatory conditions. In someembodiments, LPA-dependent conditions or diseases include those whereinan absolute or relative excess of LPA is present and/or observed.

In any of the aforementioned aspects the LPA-dependent or LPA-mediateddiseases or conditions include, but are not limited to, organ fibrosis,asthma, allergic disorders, chronic obstructive pulmonary disease,pulmonary hypertension, lung or pleural fibrosis, peritoneal fibrosis,arthritis, allergy, cancer, cardiovascular disease, ult respiratorydistress syndrome, myocardial infarction, aneurysm, stroke, and cancer.

In one aspect, compounds of Formula (I), Formula (II) and Formula (III)are used to improve the corneal sensitivity decrease caused by cornealoperations such as laser-assisted in situ keratomileusis (LASIK) orcataract operation, corneal sensitivity decrease caused by cornealdegeneration, and dry eye symptom caused thereby.

In one aspect, presented herein is the use of compounds of Formula (I),Formula (II) and Formula (III) in the treatment or prevention of ocularinflammation and allergic conjunctivitis, vernal keratoconjunctivitis,and papillary conjunctivitis in a mammal comprising administering atleast once to the mammal an effective amount of at least one compound ofFormula (I), Formula (II) or Formula (III).

In one aspect, presented herein is the use of a compound of Formula (I),Formula (II) or Formula (III) in the treatment or prevention of Sjogrendisease or inflammatory disease with dry eyes in a mammal comprisingadministering at least once to the mammal an effective amount of atleast one compound of Formula (I), Formula (II) or Formula (III).

In one aspect, LPA and LPA receptors (e.g. LPA₁) are involved in thepathogenesis of osteoarthritis (Kotani et al, Hum. Mol. Genet., 2008,17, 1790-1797). In one aspect, presented herein is the use of compoundsof Formula (I), Formula (II) and Formula (III) in the treatment orprevention of osteoarthritis in a mammal comprising administering atleast once to the mammal an effective amount of at least one compound ofFormula (I), Formula (II) or Formula (III).

In one aspect, LPA receptors (e.g. LPA₁, LPA₃) contribute to thepathogenesis of rheumatoid arthritis (Zhao et al, Mol. Pharmacol., 2008,73(2), 587-600). In one aspect, presented herein is the use of compoundsof Formula (I), Formula (II) and Formula (III) in the treatment orprevention of rheumatoid arthritis in a mammal comprising administeringat least once to the mammal an effective amount of at least one compoundof Formula (I), Formula (II) or Formula (III).

In one aspect, LPA receptors (e.g. LPA₁) contribute to adipogenesis.(Simon et al, J. Biol. Chem., 2005, vol. 280, no. 15, p.14656). In oneaspect, presented herein is the use of compounds of Formula (I), Formula(II) and Formula (III) in the promotion of adipose tissue formation in amammal comprising administering at least once to the mammal an effectiveamount of at least one compound of Formula (I), Formula (II) or Formula(III).

Compounds

Compounds of Formula (I), Formula (II) and Formula (III), includingpharmaceutically acceptable salts, prodrugs, pharmaceutically acceptablesolvates thereof, antagonize or modulate LPA receptors and are used totreat mammals diagnosed with a LPA-dependent or LPA-mediated conditionor disease. In one aspect, compounds of Formula (I), Formula (II) andFormula (III), including pharmaceutically acceptable salts, prodrugs,and pharmaceutically acceptable solvates thereof, antagonize or modulateat least one LPA receptor and are used to treat mammals suffering fromLPA-dependent or LPA-mediated conditions or diseases.

In one aspect, provided herein is a compound of Formula (I),pharmaceutically acceptable salt, pharmaceutically acceptable solvate,or prodrug thereof:

-   -   wherein,    -   R¹ is —CO₂H, —CO₂R^(A), —C(═O)NHSO₂R¹⁰, —C(═O)N(R⁹)₂,        —C(═O)NH—OH, —C(═O)NH—CN, tetrazolyl, —OH, —OR⁹, substituted or        unsubstituted monocyclic heteroaryl, or substituted or        unsubstituted monocyclic heterocycloalkyl;    -   R^(A) is H or C₁-C₆alkyl;    -   L¹ is absent, a substituted or unsubstituted alkylene, a        substituted or unsubstituted fluoroalkylene, a substituted or        unsubstituted heteroalkylene, a substituted or unsubstituted        cycloalkylene, a substituted or unsubstituted        heterocycloalkylene, a substituted or unsubstituted arylene, a        substituted or unsubstituted heteroarylene, or -L²-B-L³-;        -   L² is absent, a substituted or unsubstituted alkylene, a            substituted or unsubstituted fluoroalkylene, or a            substituted or unsubstituted heteroalkylene;        -   B is a substituted or unsubstituted cycloalkylene, a            substituted or unsubstituted heterocycloalkylene, a            substituted or unsubstituted arylene, or a substituted or            unsubstituted heteroarylene;        -   L³ is absent, a substituted or unsubstituted alkylene, a            substituted or unsubstituted fluoroalkylene, or a            substituted or unsubstituted heteroalkylene;    -   R³ is H, C₁-C₄alkyl, C₃-C₆cycloalkyl, or C₁-C₄fluoroalkyl;    -   R⁴ is —NR⁷C(═O)OCH(R⁸)—CY, —NR⁷C(═O)NR⁷CH(R⁸)—CY,        —NR⁷C(═O)(CH(R⁸))_(n)—CY, or —NR⁷C(═O)O—CY;        -   R⁷ is H or C₁-C₄alkyl;        -   R⁸ is H, C₁-C₄alkyl, or C₁-C₄fluoroalkyl;        -   CY is a substituted or unsubstituted cycloalkyl, a            substituted or unsubstituted heterocycloalkyl, a substituted            or unsubstituted aryl, or a substituted or unsubstituted            heteroaryl; or        -   R⁸ and CY are taken together with the carbon atom to which            they are attached to form a substituted or unsubstituted            carbocycle or a substituted or unsubstituted heterocycle;        -   n is 0, 1, or 2;    -   R⁵ and R⁶ are each independently selected from H, halogen, —CN,        —NO₂, —OH, —OR¹⁰, —S(═O)₂R¹⁰, —N(R⁹)S(═O)₂R¹⁰, —S(═O)₂N(R⁹)₂,        —C(═O)R¹⁰, —OC(═O)R¹⁰, —CO₂R⁹, —OCO₂R¹⁰, —N(R⁹)₂, —C(═O)N(R⁹)₂,        —OC(═O)N(R⁹)₂, —NR⁹C(═O)N(R⁹)₂, —NR⁹C(═O)R¹⁰, —NR⁹C(═O)OR¹⁰,        C₁-C₄alkyl, C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, C₁-C₄alkoxy,        and C₁-C₄heteroalkyl;    -   each R⁹ is independently selected from H, C₁-C₆alkyl,        C₁-C₆heteroalkyl, C₁-C₆fluoroalkyl, a substituted or        unsubstituted cycloalkyl, a substituted or unsubstituted        heterocycloalkyl, a substituted or unsubstituted aryl, a        substituted or unsubstituted heteroaryl, a substituted or        unsubstituted —C₁-C₄alkylene-cycloalkyl, a substituted or        unsubstituted —C₁-C₄alkylene-heterocycloalkyl, a substituted or        unsubstituted —C₁-C₄alkylene-aryl, or a substituted or        unsubstituted —C₁-C₄alkylene-heteroaryl; or    -   two R⁹ groups attached to the same N atom are taken together        with the N atom to which they are attached to form a substituted        or unsubstituted heterocycle;    -   R¹⁰ is selected from C₁-C₆alkyl, C₁-C₆heteroalkyl,        C₁-C₆fluoroalkyl, a substituted or unsubstituted cycloalkyl, a        substituted or unsubstituted heterocycloalkyl, a substituted or        unsubstituted aryl, a substituted or unsubstituted heteroaryl, a        substituted or unsubstituted —C₁-C₄alkylene-cycloalkyl, a        substituted or unsubstituted —C₁-C₄alkylene-heterocycloalkyl, a        substituted or unsubstituted —C₁-C₄alkylene-aryl, and a        substituted or unsubstituted —C₁-C₄alkylene-heteroaryl.

For any and all of the embodiments, substituents are selected from amongfrom a subset of the listed alternatives. For example, in someembodiments, R³ is H, C₁-C₄alkyl, or C₁-C₄fluoroalkyl. In otherembodiments, R³ is C₁-C₄alkyl, or C₁-C₄fluoroalkyl. In yet otherembodiments, R³ is C₁-C₄alkyl. In some embodiments, R³ is —CH₃ or—CH₂CH₃. In some embodiments, R³ is H, —CH₃, —CF₃, —CH₂CH₃, —CHCH₂, orcyclopropyl. In some embodiments, R³ is —CH₃, —CF₃, —CH₂CH₃, —CHCH₂, orcyclopropyl. In one aspect, R³ is —CH₃.

In some embodiments, R¹ is —CO₂H, —CO₂R^(A), —C(═O)N(R⁹)₂, tetrazolyl,—OH, —OR⁹, substituted or unsubstituted monocyclic heteroaryl, orsubstituted or unsubstituted monocyclic heterocycloalkyl. In someembodiments, R¹ is —CO₂H, —CO₂R^(A), tetrazolyl, —OH, —OR⁹, substitutedor unsubstituted monocyclic heteroaryl, or substituted or unsubstitutedmonocyclic heterocycloalkyl. In some embodiments, R¹ is —CO₂H,—CO₂R^(A), tetrazolyl, or a substituted or unsubstituted monocyclicheteroaryl. In some embodiments, R¹ is —CO₂H, —CO₂R^(A), tetrazolyl, ora substituted or unsubstituted monocyclic heteroaryl containing 1-Natoms.

In some embodiments, R¹ is —CO₂H or —CO₂R^(A). In some embodiments,R^(A) is H, —CH₃, or —CH₂CH₃. In one aspect, R^(A) is H. In one aspect,R¹ is —CO₂H. In some embodiments, R¹ is a substituted or unsubstitutedmonocyclic heteroaryl containing 1-N atoms.

In some embodiments, R¹ is a carboxylic acid bioisostere.

In some embodiments, R⁴ is —NR⁷C(═O)OCH(R⁸)—CY, —NR⁷C(═O)NR⁷CH(R⁸)—CY,—NR⁷C(═O)(CH(R⁸))_(n)—CY, or —NR⁷C(═O)O—CY. In other embodiments, R⁴ is—NR⁷C(═O)OCH(R⁸)—CY.

In some embodiments, R⁷ is H.

In some embodiments, R⁸ is C₁-C₄alkyl or C₁-C₄fluoroalkyl. In someembodiments, R⁸ is —CH₃ or —CF₃.

In some embodiments, R⁵ and R⁶ are each independently selected from H,halogen, —CN, —NO₂, —OH, —NH₂, —NH(C₁-C₆alkyl), —N(C₁-C₆alkyl)₂,C₁-C₄alkyl, C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, C₁-C₄alkoxy, andC₁-C₄heteroalkyl.

In other embodiments, R⁵ and R⁶ are each independently selected from H,halogen, —OH, —NH₂, —NH(CH₃), —N(CH₃)₂, —CH₃, —CF₃, —OCF₃, and —OCH₃. Insome embodiments, R⁵ and R⁶ are each independently selected from H,halogen, —OH, —CH₃, —CF₃, —OCF₃, and —OCH₃. In some embodiments, R⁵ andR⁶ are each independently selected from H, halogen, —OH, and —CH₃.

In yet other embodiments, R⁵ and R⁶ are each independently selected fromH, halogen, —CH₃, and —CF₃.

In some embodiments, R⁵ and R⁶ are each H.

In some embodiments, CY is a substituted or unsubstituted cycloalkyl, asubstituted or unsubstituted aryl, or a substituted or unsubstitutedheteroaryl.

In other embodiments, CY is a substituted or unsubstituted monocycliccycloalkyl, a substituted or unsubstituted bicyclic cycloalkyl, asubstituted or unsubstituted phenyl, a substituted or unsubstitutednapthyl, a substituted or unsubstituted monocyclic heteroaryl conatining0-4 N atoms, or a substituted or unsubstituted bicyclic heteroarylconatining 0-4 N atoms.

In yet other embodiments, CY is a substituted or unsubstitutedmonocyclic cycloalkyl, a substituted or unsubstituted phenyl, or asubstituted or unsubstituted monocyclic heteroaryl containing 0-4 Natoms.

In one aspect, CY is a substituted or unsubstituted phenyl.

In some embodiments, L¹ is absent, alkylene, fluoroalkylene,heteroalkylene, a substituted or unsubstituted cycloalkylene, asubstituted or unsubstituted heterocycloalkylene, a substituted orunsubstituted arylene, a substituted or unsubstituted heteroarylene, or-L²-B-L³-.

In some embodiments, L¹ is alkylene, fluoroalkylene, heteroalkylene, asubstituted or unsubstituted cycloalkylene, a substituted orunsubstituted heterocycloalkylene, a substituted or unsubstitutedarylene, a substituted or unsubstituted heteroarylene, or -L²-B-L³-.

In some embodiments, L¹ is a C₁-C₆alkylene, C₁-C₆fluoroalkylene,C₁-C₆heteroalkylene, a substituted or unsubstituted cycloalkylene, asubstituted or unsubstituted heterocycloalkylene, a substituted orunsubstituted arylene, a substituted or unsubstituted heteroarylene, or-L²-B-L³-.

In some embodiments, L² is absent, alkylene, fluoroalkylene, orheteroalkylene; B is a substituted or unsubstituted cycloalkylene, asubstituted or unsubstituted heterocycloalkylene, a substituted orunsubstituted arylene, or a substituted or unsubstituted heteroarylene;L³ is absent, alkylene, fluoroalkylene, or heteroalkylene.

In some embodiments, L² is abesnt, C₁-C₆alkylene, C₁-C₆fluoroalkylene,or C₁-C₆heteroalkylene; B is a substituted or unsubstitutedcycloalkylene, a substituted or unsubstituted heterocycloalkylene, asubstituted or unsubstituted arylene, or a substituted or unsubstitutedheteroarylene; L³ is absent, C₁-C₆alkylene, C₁-C₆fluoroalkylene, orC₁-C₆heteroalkylene.

In some embodiments, L² is a bond, C₁-C₆alkylene, C₁-C₆fluoroalkylene,—C₁-C₆alkylene-O—, —C₁-C₆alkylene-S—, —C₁-C₆alkylene-S(═O)—,—C₁-C₆alkylene-SO₂—, or —C₁-C₄alkylene-NR²—; B is a substituted orunsubstituted cycloalkylene, a substituted or unsubstitutedheterocycloalkylene, a substituted or unsubstituted arylene, or asubstituted or unsubstituted heteroarylene; L³ is a bond, C₁-C₆alkylene,C₁-C₆fluoroalkylene, —O—C₁-C₆alkylene-, —S—C₁-C₆alkylene-,—S(═O)—C₁-C₆alkylene-, —SO₂—C₁-C₆alkylene-, or —NR²—C₁-C₄alkylene; R² isH, C₁-C₂alkyl, or C₁-C₂fluoroalkyl.

In some embodiments, L¹ is —CH₂—, —CH(CH₃)—, —C(CH₃)₂—, —CH₂CH₂—,—CH₂CH(CH₃)—, —CH₂C(CH₃)₂—, —CH₂CH₂CH₂—, or —CH₂CH₂CH₂CH₂—. In someembodiments, L¹ is —CH₂—, —CH₂CH₂—, —CH₂CH₂CH₂—, or —CH₂CH₂CH₂CH₂—.

In some embodiments, L¹ is —CH₂—, —CH(CH₃)—, —C(CH₃)₂—, —CH₂CH₂—,—CH₂CH(CH₃)—, —CH₂C(CH₃)₂—, —CH₂CH₂CH₂—, —CH₂CH₂CH₂CH₂—, —CH₂OCH₂—,—CH₂OCH₂CH₂—, —CH₂CH₂OCH₂—, —CH₂SCH₂—, —CH₂SCH₂CH₂—, —CH₂CH₂SCH₂—,—CH₂SO₂CH₂—, —CH₂SO₂CH₂CH₂—, —CH₂CH₂SO₂CH₂—, —CH₂NHCH₂CH₂—, or—CH₂CH₂NHCH₂—. In some embodiments, L¹ is —CH₂—, —CH₂CH₂—, —CH₂CH₂CH₂—,—CH₂CH₂CH₂CH₂—, —CH₂OCH₂—, —CH₂OCH₂CH₂—, —CH₂CH₂OCH₂—, —CH₂SCH₂—,—CH₂SCH₂CH₂—, or —CH₂CH₂SCH₂—.

In some embodiments, L² is —CH₂—, —CH₂CH₂—, —CH₂O—, or —CH₂S—.

In some embodiments, L³ is —CH₂—, —CH₂CH₂—, —OCH₂—, or —SCH₂—.

In some embodiments, B is a substituted or unsubstituted monocycliccycloalkylene, a substituted or unsubstituted monocyclicheterocycloalkylene, a substituted or unsubstituted monocyclic arylene,or a substituted or unsubstituted monocyclic heteroarylene.

In some embodiments, B is a substituted or unsubstitutedC₃-C₆cycloalkylene, a substituted or unsubstitutedC₂-C₆heterocycloalkylene, a substituted or unsubstituted phenylene, asubstituted or unsubstituted naphthylene, or a substituted orunsubstituted monocyclic heteroarylene containing 0-4 N atoms or asubstituted or unsubstituted bicyclic heteroarylene containing 0-4 Natoms.

In some embodiments, B is a substituted or unsubstitutedC₃-C₆cycloalkylene, a substituted or unsubstituted phenylene, or asubstituted or unsubstituted monocyclic heteroarylene containing 0-4 Natoms.

In some embodiments, B is a substituted or unsubstituted phenylene, or asubstituted or unsubstituted monocyclic heteroarylene containing 0-4 Natoms. In some embodiments, B is a substituted or unsubstitutedphenylene, or a substituted or unsubstituted monocyclicC₁-C₅heteroarylene containing 0-4 N atoms. In some embodiments, B is asubstituted or unsubstituted phenylene, or a substituted orunsubstituted monocyclic C₁-C₅heteroarylene containing 1-4 N atoms. Insome embodiments, B is a substituted or unsubstituted phenylene, or asubstituted or unsubstituted monocyclic C₁-C₅heteroarylene containing1-3 N atoms. In some embodiments, B is a substituted or unsubstitutedphenylene, or a substituted or unsubstituted monocyclicC₁-C₅heteroarylene containing 1 or 2 N atoms.

In one aspect, in any of the embodiments described herein, B isunsubstituted or monosubstituted. In some embodiments, in any of theembodiments described herein, B is unsubstituted or monosubstituted witha group selected from halogen, C₁-C₄alkyl, C₁-C₄fluoroalkyl,C₁-C₄fluoroalkoxy, C₁-C₄alkoxy, and C₁-C₄heteroalkyl. In someembodiments, in any of the embodiments described herein, B isunsubstituted or monosubstituted with a group selected from halogen,—OH, C₁-C₄alkyl, C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, C₁-C₄alkoxy, andC₁-C₄heteroalkyl. In some embodiments, in any of the embodimentsdescribed herein, B is unsubstituted or monosubstituted with a groupselected from halogen, —OH, —CH₃, —CF₃, —OCH₃ and —OCF₃.

In one aspect, in any of the embodiments described herein, CY isunsubstituted or monosubstituted. In one aspect, in any of theembodiments described herein, CY is unsubstituted or monosubstitutedwith a group selected from halogen, C₁-C₄alkyl, C₁-C₄fluoroalkyl,C₁-C₄fluoroalkoxy, C₁-C₄alkoxy, and C₁-C₄heteroalkyl.

In some embodiments, L¹ is a C₁-C₆alkylene, C₁-C₆fluoroalkylene,C₁-C₆heteroalkylene, a substituted or unsubstituted C₃-C₆cycloalkylene,a substituted or unsubstituted C₂-C₆heterocycloalkylene, a substitutedor unsubstituted phenylene, a substituted or unsubstituted napthylene, asubstituted or unsubstituted monocyclic heteroarylene containing 0-4 Natoms or a substituted or unsubstituted bicyclic heteroarylenecontaining 0-4 N atoms, or -L²-B-L³-.

In some embodiments, L¹ is a C₁-C₆alkylene, C₁-C₆fluoroalkylene,C₁-C₆heteroalkylene, a substituted or unsubstituted C₃-C₆cycloalkylene,a substituted or unsubstituted phenylene, a substituted or unsubstitutedmonocyclic heteroarylene containing 0-4 N atoms, or -L²-B-L³-.

In some embodiments, L¹ is a C₁-C₆alkylene, C₁-C₆fluoroalkylene, orC₁-C₆heteroalkylene.

In some embodiments, L¹ is a C₁-C₆alkylene.

In some embodiments, L¹ is -L²-B-L³-.

In some embodiments, R⁴ is

In some embodiments, R⁸ is —CH₃ or —CF₃.

In one aspect, R⁸ is —CH₃.

In one aspect, R⁴ is

In another aspect, R⁴ is

In some embodiments, CY is a substituted or unsubstituted phenyl, whereif CY is substituted, then each substituent on CY is selected from H andhalogen. In one aspect, CY is 2-fluorophenyl or 2-chloro-phenyl.

In some embodiments, CY is a substituted or unsubstituted phenyl, whereif CY is substituted then CY is substituted with 1 or 2 R^(C).

In one aspect, the compound of Formula (I) has the structure of Formula(II):

In another aspect, the compound of Formula (I) has the structure ofFormula (III):

In one aspect, the compound of Formula (I) has a structure selectedfrom:

In one aspect, provided is a1-substituted-2-(4-{4-[1-(2-R^(C)-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-ethynecompound or a1-substituted-2-(3-{4-[1-(2-R^(C)-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-ethynecompound, wherein the 1-substituent is -L¹-R¹, or pharmaceuticallyacceptable salts, pharmaceutically acceptable solvates, orpharmaceutically acceptable prodrugs thereof; R¹ is —CO₂H, —CO₂R^(A),—C(═O)NHSO₂R¹⁰, —C(═O)N(R⁹)₂, —C(═O)NH—OH, —C(═O)NH—CN, tetrazolyl, —OH,—OR⁹, substituted or unsubstituted monocyclic heteroaryl, or substitutedor unsubstituted monocyclic heterocycloalkyl; R^(A) is H or C₁-C₆alkyl;L¹ is absent, a substituted or unsubstituted alkylene, a substituted orunsubstituted fluoroalkylene, a substituted or unsubstitutedheteroalkylene, a substituted or unsubstituted cycloalkylene, asubstituted or unsubstituted heterocycloalkylene, a substituted orunsubstituted arylene, a substituted or unsubstituted heteroarylene, or-L²-B-L³-; L² is absent, a substituted or unsubstituted alkylene, asubstituted or unsubstituted fluoroalkylene, or a substituted orunsubstituted heteroalkylene; B is a substituted or unsubstitutedcycloalkylene, a substituted or unsubstituted heterocycloalkylene, asubstituted or unsubstituted arylene, or a substituted or unsubstitutedheteroarylene; L³ is absent, a substituted or unsubstituted alkylene, asubstituted or unsubstituted fluoroalkylene, or a substituted orunsubstituted heteroalkylene; R^(C) is H, halogen, C₁-C₄alkyl,C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, C₁-C₄alkoxy, and C₁-C₄heteroalkyl;each R⁹ is independently selected from H, C₁-C₆alkyl, C₁-C₆heteroalkyl,C₁-C₆fluoroalkyl, a substituted or unsubstituted cycloalkyl, asubstituted or unsubstituted heterocycloalkyl, a substituted orunsubstituted aryl, a substituted or unsubstituted heteroaryl, asubstituted or unsubstituted —C₁-C₄alkylene-cycloalkyl, a substituted orunsubstituted —C₁-C₄alkylene-heterocycloalkyl, a substituted orunsubstituted —C₁-C₄alkylene-aryl, or a substituted or unsubstituted—C₁-C₄alkylene-heteroaryl; or two R⁹ groups attached to the same N atomare taken together with the N atom to which they are attached to form asubstituted or unsubstituted heterocycle; R¹⁰ is selected fromC₁-C₆alkyl, C₁-C₆heteroalkyl, C₁-C₆fluoroalkyl, a substituted orunsubstituted cycloalkyl, a substituted or unsubstitutedheterocycloalkyl, a substituted or unsubstituted aryl, a substituted orunsubstituted heteroaryl, a substituted or unsubstituted—C₁-C₄alkylene-cycloalkyl, a substituted or unsubstituted—C₁-C₄alkylene-heterocycloalkyl, a substituted or unsubstituted—C₁-C₄alkylene-aryl, and a substituted or unsubstituted—C₁-C₄alkylene-heteroaryl.

In one aspect, provided is a1-substituted-2-(4-{4-[1-(2-R^(C)-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-ethynecompound, wherein the 1-substituent is -L¹-R¹, or pharmaceuticallyacceptable salts, pharmaceutically acceptable solvates, orpharmaceutically acceptable prodrugs thereof.

In another aspect, provided is a1-substituted-2-(3-{4-[1-(2-R^(C)-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-ethynecompound, wherein the 1-substituent is -L¹-R¹, or pharmaceuticallyacceptable salts, pharmaceutically acceptable solvates, orpharmaceutically acceptable prodrugs thereof.

In some embodiments, R^(C) is H, halogen, —OH, C₁-C₄alkyl,C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, C₁-C₄alkoxy, or C₁-C₄heteroalkyl.In some embodiments, R^(C) is H, halogen, —OH, C₁-C₄alkyl,C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, or C₁-C₄alkoxy. In someembodiments, R^(C) is H, halogen, —OH, —CH₃, —CF₃, —OCF₃, or —OCH₃.

In one aspect, R^(C) is halogen. In some embodiments, R^(C) is Cl. Inother embodiments, R^(C) is F.

In one aspect, L¹ is as described in Table 1, Table 2 and/or Table 3. Inone aspect, L² is as described in Table 2. In one aspect, B is asdescribed in Table 2. In one aspect, L³ is as described in Table 2. Inone aspect, R⁸ is as described in Table 1 and/or Table 2. In one aspect,CY is as described in Table 1, Table 2 and/or Table 3. In one aspect, R¹is as described in Table 3.

Any combination of the groups described above for the various variablesis contemplated herein. Throughout the specification, groups andsubstituents thereof are chosen by one skilled in the field to providestable moieties and compounds.

In one aspect, compounds of Formula (I) include, but are not limited to,those described in Table 1, Table 2 and Table 3:

TABLE 1

Cmpd # L¹ CY R⁸ 1-1 —CH₂CH₂— 2-chlorophenyl CH₃ 1-2 —CH₂CH₂CH₂—2-chlorophenyl CH₃ 1-3 —CH₂CH₂CH₂CH₂— 2-chlorophenyl CH₃ 1-4—CH₂C(CH₃)2— 2-chlorophenyl CH₃ 1-5 —CH₂CH₂— 2-fluorophenyl CH₃ 1-6—CH₂CH₂CH₂— 2-fluorophenyl CH₃ 1-7 —CH₂CH₂CH₂CH₂— 2-fluorophenyl CH₃ 1-8—CH₂C(CH₃)₂— 2-fluorophenyl CH₃

TABLE 2

Cmpd # L² B L³ R⁸ CY 2-1  —CH₂—

— —CH₃ 2-chloro- phenyl 2-2  —CH₂—

— —CH₃ 2-chloro- phenyl 2-3  —CH₂—

— —CH₃ 2-chloro- phenyl 2-4  — phen-1,4-ylene — —CH₃ 2-chlorophenyl 2-5 — phen-1,3-ylene — —CH₃ 2-chlorophenyl 2-6  — phen-1,2-ylene — —CH₃2-chlorophenyl 2-7  — phen-1,4-ylene —CH₂— —CH₃ 2-chlorophenyl 2-8  —phen-1,3-ylene —CH₂— —CH₃ 2-chlorophenyl 2-9  — phen-1,2-ylene —CH₂——CH₃ 2-chlorophenyl 2-10 — phen-1,4-ylene —OCH₂— —CH₃ 2-chlorophenyl2-11 — phen-1,3-ylene —OCH₂— —CH₃ 2-chlorophenyl 2-12 — phen-1,2-ylene—OCH₂— —CH₃ 2-chlorophenyl 2-13 — phen-1,4-ylene —SCH₂— —CH₃2-chlorophenyl 2-14 — phen-1,3-ylene —SCH₂— —CH₃ 2-chlorophenyl 2-15 —phen-1,2-ylene —SCH₂— —CH₃ 2-chlorophenyl 2-16 —CH₂— phen-1,4-ylene ——CH₃ 2-chlorophenyl 2-17 —CH₂— phen-1,3-ylene — —CH₃ 2-chlorophenyl 2-18—CH₂— phen-1,2-ylene — —CH₃ 2-chlorophenyl 2-19 —CH₂O— phen-1,4-ylene ——CH₃ 2-chlorophenyl 2-20 —CH₂O— phen-1,3-ylene — —CH₃ 2-chlorophenyl2-21 —CH₂O— phen-1,2-ylene — —CH₃ 2-chlorophenyl 2-22 —CH₂O—phen-1,4-ylene — —CH₃ 2-chlorophenyl 2-23 —CH₂O— phen-1,3-ylene — —CH₃2-chlorophenyl 2-24 —CH₂O— phen-1,2-ylene — —CH₃ 2-chlorophenyl 2-25—CH₂S— phen-1,4-ylene — —CH₃ 2-chlorophenyl 2-26 —CH₂S— phen-1,3-ylene ——CH₃ 2-chlorophenyl 2-27 —CH₂S— phen-1,2-ylene — —CH₃ 2-chlorophenyl2-28 —CH₂— phen-1,4-ylene —CH₂— —CH₃ 2-chlorophenyl 2-29 —CH₂—phen-1,3-ylene —CH₂— —CH₃ 2-chlorophenyl 2-30 —CH₂— phen-1,2-ylene —CH₂——CH₃ 2-chlorophenyl 2-31 —CH₂O— phen-1,4-ylene —CH₂— —CH₃ 2-chlorophenyl2-32 —CH₂O— phen-1,3 -ylene —CH₂— —CH₃ 2-chlorophenyl 2-33 —CH₂O—phen-1,2-ylene —CH₂— —CH₃ 2-chlorophenyl 2-34 —CH₂O— phen-1,4-ylene—CH₂— —CH₃ 2-chlorophenyl 2-35 —CH₂O— phen-1,3 -ylene —CH₂— —CH₃2-chlorophenyl 2-36 —CH₂O— phen-1,2-ylene —CH₂— —CH₃ 2-chlorophenyl 2-37—CH₂S— phen-1,4-ylene —CH₂— —CH₃ 2-chlorophenyl 2-38 —CH₂S—phen-1,3-ylene —CH₂— —CH₃ 2-chlorophenyl 2-39 —CH₂S— phen-1,2-ylene—CH₂— —CH₃ 2-chlorophenyl 2-40 —CH₂—

— —CH₃ 2-chlorophenyl 2-41 —CH₂—

— —CH₃ 2-chlorophenyl 2-42 —CH₂—

— —CH₃ 2-chlorophenyl 2-43 — phen-1,4-ylene — —CH₃ 2-fluorophenyl 2-44 —phen-1,3-ylene — —CH₃ 2-fluorophenyl 2-45 — phen-1,2-ylene — —CH₃2-fluorophenyl 2-46 — phen-1,4-ylene —CH₂— —CH₃ 2-fluorophenyl 2-47 —phen-1,3-ylene —CH₂— —CH₃ 2-fluorophenyl 2-48 — phen-1,2-ylene —CH₂——CH₃ 2-fluorophenyl 2-49 — phen-1,4-ylene —OCH₂— —CH₃ 2-fluorophenyl2-50 — phen-1,3-ylene —OCH₂— —CH₃ 2-fluorophenyl 2-51 — phen-1,2-ylene—OCH₂— —CH₃ 2-fluorophenyl 2-52 — phen-1,4-ylene —SCH₂— —CH₃2-fluorophenyl 2-53 — phen-1,3-ylene —SCH₂— —CH₃ 2-fluorophenyl 2-54 —phen-1,2-ylene —SCH₂— —CH₃ 2-fluorophenyl 2-55 —CH₂— phen-1,4-ylene ——CH₃ 2-fluorophenyl 2-56 —CH₂— phen-1,3-ylene — —CH₃ 2-fluorophenyl 2-57—CH₂— phen-1,2-ylene — —CH₃ 2-fluorophenyl 2-58 —CH₂O— phen-1,4-ylene ——CH₃ 2-fluorophenyl 2-59 —CH₂O— phen-1,3-ylene — —CH₃ 2-fluorophenyl2-60 —CH₂O— phen-1,2-ylene — —CH₃ 2-fluorophenyl 2-61 —CH₂S—phen-1,4-ylene — —CH₃ 2-fluorophenyl 2-62 —CH₂S— phen-1,3-ylene — —CH₃2-fluorophenyl 2-63 —CH₂S— phen-1,2-ylene — —CH₃ 2-fluorophenyl 2-64—CH₂— phen-1,4-ylene —CH₂— —CH₃ 2-fluorophenyl 2-65 —CH₂— phen-1,3-ylene—CH₂— —CH₃ 2-fluorophenyl 2-66 —CH₂— phen-1,2-ylene —CH₂— —CH₃2-fluorophenyl 2-67 —CH₂O— phen-1,4-ylene —CH₂— —CH₃ 2-fluorophenyl 2-68—CH₂O— phen-1,3-ylene —CH₂— —CH₃ 2-fluorophenyl 2-69 —CH₂O—phen-1,2-ylene —CH₂— —CH₃ 2-fluorophenyl 2-70 —CH₂S— phen-1,4-ylene—CH₂— —CH₃ 2-fluorophenyl 2-71 —CH₂S— phen-1,3-ylene —CH₂— —CH₃2-fluorophenyl 2-72 —CH₂S— phen-1,2-ylene —CH₂— —CH₃ 2-fluorophenyl

TABLE 3

Cmpd # L¹ CY R¹ 3-1 —CH₂— 2-chlorophenyl 1,2,4-triazol-1-yl 3-2 —CH₂CH₂—2-chlorophenyl 1,2,4-triazol-1-yl 3-3 —CH₂— 2-chlorophenyl thiazol-2-yl3-4 —CH₂— 2-chlorophenyl pyrimidin-2-yl 3-5 —CH₂— 2-fluorophenyl1,2,4-triazol-1-yl 3-6 —CH₂CH₂— 2-fluorophenyl 1,2,4-triazol-1-yl 3-7—CH₂— 2-fluorophenyl thiazol-2-yl 3-8 —CH₂— 2-fluorophenylpyrimidin-2-ylSynthesis of Compounds

Compounds of Formula (I), Formula (II) and Formula (III) describedherein are synthesized using standard synthetic techniques or usingmethods known in the art in combination with methods described herein.In additions, solvents, temperatures and other reaction conditionspresented herein may vary.

The starting material used for the synthesis of the compounds of Formula(I), Formula (II) and Formula (III) are either synthesized or obtainedfrom commercial sources, such as, but not limited to, Sigma-Aldrich,Fluka, Acros Organics, Alfa Aesar, and the like. The compounds describedherein, and other related compounds having different substituents aresynthesized using techniques and materials described herein or otherwiseknown, including those found in March, ADVANCED ORGANIC CHEMISTRY 4^(th)Ed., (Wiley 1992); Carey and Sundberg, ADVANCED ORGANIC CHEMISTRY 4^(th)Ed., Vols. A and B (Plenum 2000, 2001), and Green and Wuts, PROTECTIVEGROUPS IN ORGANIC SYNTHESIS 3^(rd) Ed., (Wiley 1999). General methodsfor the preparation of compounds can be modified by the use ofappropriate reagents and conditions for the introduction of the variousmoieties found in the formulae as provided herein.

In some embodiments, the compounds of Formula (I), Formula (II) andFormula (III) are prepared as outlined in the following Schemes.

The synthesis of compounds of Formula (I), Formula (II) and Formula(III) described herein begins with the reaction of an alkyl acetoacetatewith methylamine to provide a compound of structure II. Compounds ofstructure II are reacted with a substituted or unsubstituted4-halo-benzoyl chloride (structure III) to provide compounds ofstructure IV. The formation of isoxazoles of structure V from compoundsof structure IV proceeds in the presence of hydroxyl amine and aceticacid. Hydrolysis of the ester of isoxazoles of structure V providescarboxylic acids of structure VI. A Curtius rearrangement of carboxylicacids of structure VI in the presence of hydroxy compounds of structureVII provides carbamate compounds of structure VIII.

In one aspect, compounds of structure VIII are used in the synthesis ofcompounds of Formula (I), Formula (II) and Formula (III) as shown inScheme 2.

Reaction of compounds of structure VIII with alkyne of structure IXunder Sonagashira reaction conditions provides alkynes of structure X.Hydrolysis of the ester group of alkynes of structure X providescompounds of structure XI.

Sonagahira reaction conditions include those described herein and thoseknown in the art. Examplary Sonagashira reaction conditions includethose described in K. Sonogashira et al., Tetrahedron Letters 16 (50):4467-4470, 1975; Rafael Chinchilla et al., Chem. Rev.; 107(3) pp874-922, 2007; Gil-Molto et al., Advanced Synthesis & Catalysis,348(14), 1874-1882, 2006; Eckhardt, G. C. Fu, J. Am. Chem. Soc., 125,13642-13643, 2003; Y. Liang et al., J. Org. Chem., 71, 379-381, 2006; P.Li et al., Tetrahedron, 61, 8633-8640, 2005; A. R. Gholap et al., J.Org. Chem., 2005, 70, 4869-4872; C. Yi, R. Hua, J. Org. Chem., 71,2535-2537, 2006; D. Gelman, S. L. Buchwald, Angew. Chem. Int. Ed., 2003,42, 5993-5996; B. Liang et al., J. Org. Chem., 70, 391-393, 2005; A.Elangovan et al., Org. Lett., 2003, 5, 1841-1844; Batey et al., Org.Lett., 1411-1414, 2002; Lemay et al., J. Org. Chem., 71, 3615-3618,2006; A. S. Karpov, T. J. J. Muller, Synthesis, 2003, 2815-2826.

In one aspect, compounds of sturcture VIII are used in the synthesis ofcompounds of Formula (I), Formula (II) and Formula (III) in thestep-wise manner shown Scheme 3.

Reaction of propargyl alcohol with compounds of structure VIII underSonagashira reaction conditions affords compounds of structure XII.Activation of the propargyl alcohol of structure XII followed bydisplacement with a nucleophile of structure XIII addords compounds ofstructure XV. In one aspect, activation of the propargyl alcohol ofcompounds of structure XII is achieved with triphenyl phosphine. Inother embodiments, the propargyl alcohol is activated and reacted withcompounds of structure XIV to provide compounds of structure XVI. In oneaspect, when Ra is alkyl, compounds of structure XV and XVI arehydrolyzed to provide the carboxylic acids.

In one aspect, compounds of Formula (I), Formula (II) and Formula (III)are synthesized as outlined in the Examples.

Throughout the specification, groups and substituents thereof are chosenby one skilled in the field to provide stable moieties and compounds.

A detailed description of techniques applicable to the creation ofprotecting groups and their removal are described in Greene and Wuts,Protective Groups in Organic Synthesis, 3rd Ed., John Wiley & Sons, NewYork, N.Y., 1999, and Kocienski, Protective Groups, Thieme Verlag, NewYork, N.Y., 1994, which are incorporated herein by reference for suchdisclosure.

Further Forms of Compounds

In one aspect, compounds of Formula (I), Formula (II) and Formula (III)possess one or more stereocenters and each stereocenter existsindependently in either the R or S configuration. The compoundspresented herein include all diastereomeric, enantiomeric, and epimericforms as well as the appropriate mixtures thereof. The compounds andmethods provided herein include all cis, trans, syn, anti, entgegen (E),and zusammen (Z) isomers as well as the appropriate mixtures thereof. Incertain embodiments, compounds of Formula (I), Formula (II) and Formula(III) are prepared as their individual stereoisomers by reacting aracemic mixture of the compound with an optically active resolving agentto form a pair of diastereoisomeric compounds/salts, separating thediastereomers and recovering the optically pure enantiomers. In someembodiments, resolution of enantiomers is carried out using covalentdiastereomeric derivatives of the compounds described herein. In anotherembodiment, diastereomers are separated by separation/resolutiontechniques based upon differences in solubility. In other embodiments,separation of steroisomers is performed by chromatography or by theforming diastereomeric salts and separation by recrystallization, orchromatography, or any combination thereof. Jean Jacques, Andre Collet,Samuel H. Wilen, “Enantiomers, Racemates and Resolutions”, John WileyAnd Sons, Inc., 1981. In one aspect, stereoisomers are obtained bystereoselective synthesis.

The methods and compositions described herein include the use ofamorphous forms as well as crystalline forms (also known as polymorphs).In one aspect, compounds described herein are in the form ofpharmaceutically acceptable salts. As well, active metabolites of thesecompounds having the same type of activity are included in the scope ofthe present disclosure. In addition, the compounds described herein canexist in unsolvated as well as solvated forms with pharmaceuticallyacceptable solvents such as water, ethanol, and the like. The solvatedforms of the compounds presented herein are also considered to bedisclosed herein.

In some embodiments, compounds described herein are prepared asprodrugs. A “prodrug” refers to an agent that is converted into theparent drug in vivo. Prodrugs are often useful because, in somesituations, they may be easier to administer than the parent drug. Theymay, for instance, be bioavailable by oral administration whereas theparent is not. The prodrug may also have improved solubility inpharmaceutical compositions over the parent drug. In some embodiments,the design of a prodrug increases the effective water solubility. Anexample, without limitation, of a prodrug is a compound describedherein, which is administered as an ester (the “prodrug”) to facilitatetransmittal across a cell membrane where water solubility is detrimentalto mobility but which then is metabolically hydrolyzed to the carboxylicacid, the active entity, once inside the cell where water-solubility isbeneficial. A further example of a prodrug might be a short peptide(polyaminoacid) bonded to an acid group where the peptide is metabolizedto reveal the active moiety. In certain embodiments, upon in vivoadministration, a prodrug is chemically converted to the biologically,pharmaceutically or therapeutically active form of the compound. Incertain embodiments, a prodrug is enzymatically metabolized by one ormore steps or processes to the biologically, pharmaceutically ortherapeutically active form of the compound.

In one aspect, prodrugs are designed to alter the metabolic stability orthe transport characteristics of a drug, to mask side effects ortoxicity, to improve the flavor of a drug or to alter othercharacteristics or properties of a drug. By virtue of knowledge ofpharmacokinetic, pharmacodynamic processes and drug metabolism in vivo,once a pharmaceutically active compound is known, the design prodrugs ofthe compound is possible. (see, for example, Nogrady (1985) MedicinalChemistry A Biochemical Approach, Oxford University Press, New York,pages 388-392; Silverman (1992), The Organic Chemistry of Drug Designand Drug Action, Academic Press, Inc., San Diego, pages 352-401,Rooseboom et al., Pharmacological Reviews, 56:53-102, 2004; Aesop Cho,“Recent Advances in Oral Prodrug Discovery”, Annual Reports in MedicinalChemistry, Vol. 41, 395-407, 2006; T. Higuchi and V. Stella, Pro-drugsas Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series).

Prodrug forms of the herein described compounds, wherein the prodrug ismetabolized in vivo to produce a compound of Formula (I), Formula (II)or Formula (III) as set forth herein are included within the scope ofthe claims. In some cases, some of the herein-described compounds may bea prodrug for another derivative or active compound.

In some embodiments, sites on the aromatic ring portion of compounds ofFormula (I), Formula (II) and Formula (III) are susceptible to variousmetabolic reactions Therefore incorporation of appropriate substituentson the aromatic ring structures will reduce, minimize or eliminate thismetabolic pathway. In specific embodiments, the appropriate substituentto decrease or eliminate the susceptibility of the aromatic ring tometabolic reactions is, by way of example only, a halogen, or an alkylgroup.

In another embodiment, the compounds described herein are labeledisotopically (e.g. with a radioisotope) or by another other means,including, but not limited to, the use of chromophores or fluorescentmoieties, bioluminescent labels, or chemiluminescent labels.

Compounds described herein include isotopically-labeled compounds, whichare identical to those recited in the various formulae and structurespresented herein, but for the fact that one or more atoms are replacedby an atom having an atomic mass or mass number different from theatomic mass or mass number usually found in nature. Examples of isotopesthat can be incorporated into the present compounds include isotopes ofhydrogen, carbon, nitrogen, oxygen, fluorine and chlorine, such as, forexample, ²H, 3H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³⁵S, ¹⁸F, ³⁶Cl. In one aspect,isotopically-labeled compounds described herein, for example those intowhich radioactive isotopes such as ³H and ¹⁴C are incorporated, areuseful in drug and/or substrate tissue distribution assays. In oneaspect, substitution with isotopes such as deuterium affords certaintherapeutic advantages resulting from greater metabolic stability, suchas, for example, increased in vivo half-life or reduced dosagerequirements.

In additional or further embodiments, the compounds described herein aremetabolized upon administration to an organism in need to produce ametabolite that is then used to produce a desired effect, including adesired therapeutic effect.

“Pharmaceutically acceptable,” as used herein, refers a material, suchas a carrier or diluent, which does not abrogate the biological activityor properties of the compound, and is relatively nontoxic, i.e., thematerial may be administered to an individual without causingundesirable biological effects or interacting in a deleterious mannerwith any of the components of the composition in which it is contained.

The term “pharmaceutically acceptable salt” refers to a formulation of acompound that does not cause significant irritation to an organism towhich it is administered and does not abrogate the biological activityand properties of the compound. In some embodiments, pharmaceuticallyacceptable salts are obtained by reacting a compound of Formula (I),Formula (II) or Formula (III) with acids. Pharmaceutically acceptablesalts are also obtained by reacting a compound of Formula (I), Formula(II) or Formula (III) with a base to form a salt.

Compounds described herein may be formed as, and/or used as,pharmaceutically acceptable salts. The type of pharmaceutical acceptablesalts, include, but are not limited to: (1) acid addition salts, formedby reacting the free base form of the compound with a pharmaceuticallyacceptable: inorganic acid, such as, for example, hydrochloric acid,hydrobromic acid, sulfuric acid, phosphoric acid, metaphosphoric acid,and the like; or with an organic acid, such as, for example, aceticacid, propionic acid, hexanoic acid, cyclopentanepropionic acid,glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid,malic acid, maleic acid, fumaric acid, trifluoroacetic acid, tartaricacid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid,cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonicacid, toluenesulfonic acid, 2-naphthalenesulfonic acid,4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid,4,4′-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionicacid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuricacid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylicacid, stearic acid, muconic acid, butyric acid, phenylacetic acid,phenylbutyric acid, valproic acid, and the like; (2) salts formed whenan acidic proton present in the parent compound is replaced by a metalion, e.g., an alkali metal ion (e.g. lithium, sodium, potassium), analkaline earth ion (e.g. magnesium, or calcium), or an aluminum ion. Insome cases, compounds described herein may coordinate with an organicbase, such as, but not limited to, ethanolamine, diethanolamine,triethanolamine, tromethamine, N-methylglucamine, dicyclohexylamine,tris(hydroxymethyl)methylamine. In other cases, compounds describedherein may form salts with amino acids such as, but not limited to,arginine, lysine, and the like. Acceptable inorganic bases used to formsalts with compounds that include an acidic proton, include, but are notlimited to, aluminum hydroxide, calcium hydroxide, potassium hydroxide,sodium carbonate, sodium hydroxide, and the like.

It should be understood that a reference to a pharmaceuticallyacceptable salt includes the solvent addition forms or crystal formsthereof, particularly solvates or polymorphs. Solvates contain eitherstoichiometric or non-stoichiometric amounts of a solvent, and may beformed during the process of crystallization with pharmaceuticallyacceptable solvents such as water, ethanol, and the like. Hydrates areformed when the solvent is water, or alcoholates are formed when thesolvent is alcohol. Solvates of compounds described herein can beconveniently prepared or formed during the processes described herein.In addition, the compounds provided herein can exist in unsolvated aswell as solvated forms. In general, the solvated forms are consideredequivalent to the unsolvated forms for the purposes of the compounds andmethods provided herein.

Compounds described herein, such as compounds of Formula (I), Formula(II) and Formula (III), may be in various forms, including but notlimited to, amorphous forms, milled forms and nano-particulate forms. Inaddition, compounds described herein include crystalline forms, alsoknown as polymorphs. Polymorphs include the different crystal packingarrangements of the same elemental composition of a compound. Polymorphsusually have different X-ray diffraction patterns, melting points,density, hardness, crystal shape, optical properties, stability, andsolubility. Various factors such as the recrystallization solvent, rateof crystallization, and storage temperature may cause a single crystalform to dominate.

Throughout the specification, groups and substituents thereof can bechosen by one skilled in the field to provide stable moieties andcompounds.

Certain Terminology

Unless otherwise stated, the following terms used in this application,including the specification and claims, have the definitions givenbelow. It must be noted that, as used in the specification and theappended claims, the singular forms “a,” “an” and “the” include pluralreferents unless the context clearly dictates otherwise. Unlessotherwise indicated, conventional methods of mass spectroscopy, NMR,HPLC, protein chemistry, biochemistry, recombinant DNA techniques andpharmacology are employed. In this application, the use of “or” or “and”means “and/or” unless stated otherwise. Furthermore, use of the term“including” as well as other forms, such as “include”, “includes,” and“included,” is not limiting. The section headings used herein are fororganizational purposes only and are not to be construed as limiting thesubject matter described.

An “alkyl” group refers to an aliphatic hydrocarbon group. The alkylgroup may be a saturated alkyl group (which means that it does notcontain any carbon-carbon double bonds or carbon-carbon triple bonds) orthe the alkyl group may be an unsaturated alkyl group (which means thatit contains at least one carbon-carbon double bonds or carbon-carbontriple bond). The alkyl moiety, whether saturated or unsaturated, may bebranched, or straight chain.

The “alkyl” group may have 1 to 10 carbon atoms (whenever it appearsherein, a numerical range such as “1 to 10” refers to each integer inthe given range; e.g., “1 to 10 carbon atoms” means that the alkyl groupmay consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., upto and including 10 carbon atoms, although the present definition alsocovers the occurrence of the term “alkyl” where no numerical range isdesignated). The alkyl group of the compounds described herein may bedesignated as “C₁-C₆ alkyl” or similar designations. By way of exampleonly, “C₁-C₆ alkyl” indicates that there are one, two, three, four,five, or six carbon atoms in the alkyl chain. In one aspect the alkyl isselected from the group consisting of methyl, ethyl, propyl, iso-propyl,n-butyl, iso-butyl, sec-butyl, and t-butyl. Typical alkyl groupsinclude, but are in no way limited to, methyl, ethyl, propyl, isopropyl,butyl, isobutyl, sec-butyl, tertiary butyl, pentyl, neopentyl, hexyl,allyl, but-2-enyl, but-3-enyl, and the like. In one aspect, an alkyl isa C₁-C₆ alkyl.

The term “alkylene” refers to a divalent alkyl radical. Any of the abovementioned monovalent alkyl groups may be an alkylene by abstraction of asecond hydrogen atom from the alkyl. In one aspect, an alkelene is aC₁-C₆alkylene. In another aspect, an alkylene is a C₁-C₄alkylene.Typical alkylene groups include, but are not limited to, —CH₂—,—CH(CH₃)—, —C(CH₃)₂—, —CH₂CH₂—, —CH₂CH(CH₃)—, —CH₂C(CH₃)₂—, —CH₂CH₂CH₂—,—CH₂CH₂CH₂CH₂—, and the like.

An “alkoxy” group refers to a (alkyl)O— group, where alkyl is as definedherein.

The term “alkylamine” refers to the —N(alkyl)_(x)H_(y) group, where xand y are selected from the group x=1, y=1 and x=2, y=0. In someembodiments, when x=2 and y=0, the alkyl groups taken together with thenitrogen atom to which they are attached form a cyclic ring system.

The term “aromatic” refers to a planar ring having a delocalizedπ-electron system containing 4n+2π electrons, where n is an integer.Aromatic rings can be formed from five, six, seven, eight, nine, ten, ormore than ten atoms. Aromatics are optionally substituted. The term“aromatic” includes both carbocyclic aryl (“aryl”, e.g., phenyl) andheterocyclic aryl (or “heteroaryl” or “heteroaromatic”) groups (e.g.,pyridine). The term includes monocyclic or fused-ring polycyclic (i.e.,rings which share adjacent pairs of carbon atoms) groups.

The term “carbocyclic” or “carbocycle” refers to a ring or ring systemwhere the atoms forming the backbone of the ring are all carbon atoms.The term thus distinguishes carbocyclic from heterocyclic rings in whichthe ring backbone contains at least one atom which is different fromcarbon.

As used herein, the term “aryl” refers to an aromatic ring wherein eachof the atoms forming the ring is a carbon atom. Aryl rings are formed byfive, six, seven, eight, nine, or more than nine carbon atoms. Arylgroups are optionally substituted. In one aspect, an aryl is a phenyl ora naphthalenyl. In one aspect, an aryl is a phenyl. In one aspect, anaryl is a C₆-C₁₀aryl. Depending on the structure, an aryl group can be amonoradical or a diradical (i.e., an arylene group). In one aspect, anarylene is a C₆-C₁₀ arylene. Examplary arylenes include, but are notlimited to, phenyl-1,2-ene, phenyl-1,3-ene, and phenyl-1,4-ene.

The term “cycloalkyl” refers to a monocyclic or polycyclic aliphatic,non-aromatic radical, wherein each of the atoms forming the ring (i.e.skeletal atoms) is a carbon atom. Cycloalkyls may be saturated, orpartially unsaturated. Cycloalkyls may be fused with an aromatic ring,and the point of attachment is at a carbon that is not an aromatic ringcarbon atom. Cycloalkyl groups include groups having from 3 to 10 ringatoms. Illustrative examples of cycloalkyl groups include, but are notlimited to, the following moieties:

and the like. In some embodiments, cycloalkyl groups are selected fromamong cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl,cyclohexenyl, cycloheptyl, and cyclooctyl. Cycloalkyl groups may besubstituted or unsubstituted. Depending on the structure, a cycloalkylgroup can be a monoradical or a diradical (i.e., an cycloalkylene group,such as, but not limited to, cyclopropan-1,1-diyl, cyclobutan-1,1-diyl,cyclopentan-1,1-diyl, cyclohexan-1,1-diyl, cyclohexan-1,4-diyl,cycloheptan-1,1-diyl, and the like). In one aspect, cycloalkyl is aC₃-C₆cycloalkyl.

The term “halo” or, alternatively, “halogen” or “halide” means fluoro,chloro, bromo or iodo.

The term “haloalkyl” refers to an alkyl group in which one or morehydrogen atoms are replaced by one or more halide atoms. In one aspect,a haloalkyl is a C₁-C₄haloalkyl.

The term “haloalkylene” refers to an alkylene group in which one or morehydrogen atoms are replaced by one or more halide atoms. In one aspect,a haloalkylene is a C₁-C₆haloalkylene. In another aspect, a haloalkyleneis a C₁-C₄haloalkylene.

The term “fluoroalkyl” refers to an alkyl in which one or more hydrogenatoms are replaced by a fluorine atom. In one aspect, a fluoralkyl is aC₁-C₄fluoroalkyl.

The term “fluoroalkylene” refers to an alkylene in which one or morehydrogen atoms are replaced by a fluorine atom. In one aspect, afluoralkylene is a C₁-C₆fluoroalkylene. In another aspect, afluoralkylene is a C₁-C₄fluoroalkylene.

The term “heteroalkyl” refers to an alkyl group in which one or moreskeletal atoms of the alkyl are selected from an atom other than carbon,e.g., oxygen, nitrogen (e.g. NH or N-alkyl), sulfur, or combinationsthereof. In one aspect, a heteroalkyl is a C₁-C₆heteroalkyl.

The term “heteroalkylene” refers to an alkylene group in which one ormore skeletal atoms of the alkyl are selected from an atom other thancarbon, e.g., oxygen, nitrogen, sulfur, phosphorus or combinationsthereof. In one aspect, a heteroalkylene is a C₁-C₆heteroalkylene. Inanother aspect, a heteroalkylene is a C₁-C₄heteroalkylene. Examplaryheteroalkylenes include, but are not limited to, —OCH₂—, —OCH(CH₃)—,—OC(CH₃)₂—, —OCH₂CH₂—, —CH₂O—, —CH(CH₃)O—, —C(CH₃)₂O—, —CH₂CH₂O—,—CH₂OCH₂—, —CH₂OCH₂CH₂—, —CH₂CH₂OCH₂—, —SCH₂—, —SCH(CH₃)—, —SC(CH₃)₂—,—SCH₂CH₂—, —CH₂S—, —CH(CH₃)S—,—C(CH₃)₂S—, —CH₂CH₂S—, —CH₂SCH₂—,—CH₂SCH₂CH₂—, —CH₂CH₂SCH₂—, —SO₂CH₂—, —SO₂CH(CH₃)—, —SO₂C(CH₃)₂—,—SO₂CH₂CH₂—, —CH₂SO₂—, —CH(CH₃)SO₂—, —C(CH₃)₂SO₂—, —CH₂CH₂SO₂—,—CH₂SO₂CH₂—, —CH₂SO₂CH₂CH₂—, —CH₂CH₂SO₂CH₂—, —NHCH₂—, —NHCH(CH₃)—,—NHC(CH₃)₂—, —NHCH₂CH₂—, —CH₂NH—, —CH(CH₃)NH—, —C(CH₃)₂NH—, —CH₂CH₂NH—,—CH₂NHCH₂—, —CH₂NHCH₂CH₂—, —CH₂CH₂NHCH₂—, and the like.

The term “heterocycle” or “heterocyclic” refers to heteroaromatic rings(also known as heteroaryls) and heterocycloalkyl rings (also known asheteroalicyclic groups) containing one to four heteroatoms in thering(s), where each heteroatom in the ring(s) is selected from O, S andN, wherein each heterocyclic group has from 4 to 10 atoms in its ringsystem, and with the proviso that the any ring does not contain twoadjacent O or S atoms. Non-aromatic heterocyclic groups (also known asheterocycloalkyls) include groups having only 3 atoms in their ringsystem, but aromatic heterocyclic groups must have at least 5 atoms intheir ring system. The heterocyclic groups include benzo-fused ringsystems. An example of a 3-membered heterocyclic group is aziridinyl. Anexample of a 4-membered heterocyclic group is azetidinyl. An example ofa 5-membered heterocyclic group is thiazolyl. An example of a 6-memberedheterocyclic group is pyridyl, and an example of a 10-memberedheterocyclic group is quinolinyl. Examples of non-aromatic heterocyclicgroups are pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl,tetrahydrothienyl, oxazolidinonyl, tetrahydropyranyl, dihydropyranyl,tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl,thioxanyl, piperazinyl, aziridinyl, azetidinyl, oxetanyl, thietanyl,homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl,thiazepinyl, 1,2,3,6-tetrahydropyridinyl, pyrrolin-2-yl, pyrrolin-3-yl,indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl,pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl,dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl,3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, 3H-indolyl andquinolizinyl. Examples of aromatic heterocyclic groups are pyridinyl,imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl,furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl,quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl,cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl,triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl,furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl,benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, andfuropyridinyl. The foregoing groups may be C-attached or N-attachedwhere such is possible. For instance, a group derived from pyrrole maybe pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached). Further, agroup derived from imidazole may be imidazol-1-yl or imidazol-3-yl (bothN-attached) or imidazol-2-yl, imidazol-4-yl or imidazol-5-yl (allC-attached). The heterocyclic groups include benzo-fused ring systems.Non-aromatic heterocycles may ber substituted with one or two oxo (═O)moieties, such as pyrrolidin-2-one.

The terms “heteroaryl” or, alternatively, “heteroaromatic” refers to anaryl group that includes one or more ring heteroatoms selected fromnitrogen, oxygen and sulfur. Illustrative examples of heteroaryl groupsinclude the following moieties:

and the like. Monocyclic heteroaryls include pyridinyl, imidazolyl,pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl,thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl,pyridazinyl, triazinyl, oxadiazolyl, thiadiazolyl, and furazanyl. In oneaspect, a heteroaryl contains 0-3 N atoms. In another aspect, aheteroaryl contains 1-3 N atoms. In another aspect, a heteroarylcontains 0-3 N atoms, 0-1 O atoms, and 0-1 S atoms. In another aspect, aheteroaryl is a monocyclic or bicyclic heteroaryl. In one aspect,heteroaryl is a C₁-C₉heteroaryl. In one aspect, monocyclic heteroaryl isa C₁-C₅heteroaryl. In one aspect, monocyclic heteroaryl is a 5-memberedor 6-membered heteroaryl. In one aspect, bicyclic heteroaryl is aC₆-C₉heteroaryl. Depending on the structure, a heteroaryl group can be amonoradical or a diradical (i.e., a heteroarylene group).

A “heterocycloalkyl” or “heteroalicyclic” group refers to a cycloalkylgroup that includes at least one heteroatom selected from nitrogen,oxygen and sulfur. The radicals may be fused with an aryl or heteroaryl.Illustrative examples of heterocycloalkyl groups, also referred to asnon-aromatic heterocycles, include:

and the like. In some embodiments, the heterocycloalkyl is selected fromoxazolidinonyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl,tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl,thiomorpholinyl, piperazinyl, and indolinyl. The term heteroalicyclicalso includes all ring forms of the carbohydrates, including but notlimited to the monosaccharides, the disaccharides and theoligosaccharides. In one aspect, a heterocycloalkyl is aC₂-C₁₀heterocycloalkyl. In another aspect, a heterocycloalkyl is aC₄-C₁₀heterocycloalkyl. In one aspect, a heterocycloalkyl contains 0-2 Natoms. In another aspect, a heterocycloalkyl contains 0-2 N atoms, 0-2 Oatoms or 0-1 S atoms.

The term “bond” or “single bond” refers to a chemical bond between twoatoms, or two moieties when the atoms joined by the bond are consideredto be part of larger substructure. In one aspect, when a group describedherein is a bond, the referenced group is absent thereby allowing a bondto be formed between the remaining identified groups.

The term “membered ring” includes any cyclic structure. The term“membered” is meant to denote the number of skeletal atoms thatconstitute the ring. Thus, for example, cyclohexyl, pyridinyl, pyranyland thiopyranyl are 6-membered rings and cyclopentyl, pyrrolyl, furanyl,and thienyl are 5-membered rings.

The term “moiety” refers to a specific segment or functional group of amolecule. Chemical moieties are often recognized chemical entitiesembedded in or appended to a molecule.

As used herein, “carboxylic acid bioisostere” refers to a functionalgroup or moiety that exhibits similar physical, biological and/orchemical properties as a carboxylic acid moiety. Examples of carboxylicacid bioisosteres include, but are not limited to,

and the like.

The term “optionally substituted” or “substituted” means that thereferenced group may be substituted with one or more additional group(s)individually and independently selected from alkyl, cycloalkyl, aryl,heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, alkylthio,arylthio, alkylsulfoxide, arylsulfoxide, alkylsulfone, arylsulfone,cyano, halo, nitro, haloalkyl, fluoroalkyl, fluoroalkoxy, and amino,including mono- and di-substituted amino groups, and the protectedderivatives thereof By way of example an optional substituents may behalide, —CN, —NO₂, or L_(s)R_(s), wherein each L_(s) is independentlyselected from a bond, —O—, —C(═O)—, —C(═O)O—, —S—, —S(═O)—, —S(═O)₂—,—NH—, —NHC(═O)—, —C(═O)NH—, S(═O)₂NH—, —NHS(═O)₂, —OC(═O)NH—,—NHC(═O)O—, or —(C₁-C₆ alkylene)-; and each R_(s) is selected from H,alkyl, fluoroalkyl, heteroalkyl, cycloalkyl, aryl, heteroaryl, orheterocycloalkyl. The protecting groups that may form the protectivederivatives of the above substituents may be found in sources such asGreene and Wuts, above. In some embodiments, optional substituents areselected from halogen, —CN, —NH₂, —OH, —N(CH₃)₂, alkyl, fluoroalkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, alkoxy,aryloxy, alkylthio, arylthio, alkylsulfoxide, arylsulfoxide,alkylsulfone, and arylsulfone. In some embodiments, an optionalsubstituents is halogen, —CN, —NH₂, —OH, —NH(CH₃), —N(CH₃)₂, alkyl,fluoroalkyl, heteroalkyl, alkoxy, fluoroalkoxy, —S-alkyl, or—S(═O)₂alkyl. In some embodiments, an optional substituent is selectedfrom halogen, —CN, —NH₂, —OH, —NH(CH₃), —N(CH₃)₂, —CH₃, —CH₂CH₃, —CF₃,—OCH₃, and —OCF₃. In some embodiments, substituted groups aresubstituted with one or two of the preceding groups. In someembodiments, substituted groups are substituted with one of thepreceding groups. In some embodiments, an optional substituent on analiphatic carbon atom (acyclic or cyclic, saturated or unsaturatedcarbon atoms, excluding aromatic carbon atoms) includes oxo (═O).

As used herein,“1-substituted-2-(4-{4-[1-(2-R^(C)-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-ethynecompound” refers to the following structure:

where the point of attachment of the 1-substitutent is denoted by

.

As used herein,“1-substituted-2-(3-{4-[1-(2-R^(C)-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-ethynecompound” refers to the following structure:

where the point of attachment of the 1-substitutent is denoted by

.

In certain embodiments, the compounds presented herein possess one ormore stereocenters and each center independently exists in either the Ror S configuration. The compounds presented herein include alldiastereomeric, enantiomeric, and epimeric forms as well as theappropriate mixtures thereof. Stereoisomers are obtained, if desired, bymethods such as, stereoselective synthesis and/or the separation ofstereoisomers by chiral chromatographic columns.

The methods and formulations described herein include the use ofN-oxides (if appropriate), crystalline forms (also known as polymorphs),or pharmaceutically acceptable salts of compounds having the structureof Formula (I), Formula (II) or Formula (III), as well as activemetabolites of these compounds having the same type of activity. In somesituations, compounds may exist as tautomers. All tautomers are includedwithin the scope of the compounds presented herein. In specificembodiments, the compounds described herein exist in solvated forms withpharmaceutically acceptable solvents such as water, ethanol, and thelike. In other embodiments, the compounds described herein exist inunsolvated form.

Certain Terminology

The term “acceptable” with respect to a formulation, composition oringredient, as used herein, means having no persistent detrimentaleffect on the general health of the subject being treated.

The term “modulate,” as used herein, means to interact with a targeteither directly or indirectly so as to alter the activity of the target,including, by way of example only, to enhance the activity of thetarget, to inhibit the activity of the target, to limit the activity ofthe target, or to extend the activity of the target.

The term “modulator,” as used herein, refers to a molecule thatinteracts with a target either directly or indirectly. The interactionsinclude, but are not limited to, the interactions of an agonist, partialagonist, an inverse agonist and antagonist. In one embodiment, amodulator is an antagonist.

The term “agonist,” as used herein, refers to a molecule such as acompound, a drug, an enzyme activator or a hormone modulator that bindsto a specific receptor and triggers a response in the cell. An agonistmimics the action of an endogenous ligand (such as LPA, prostaglandin,hormone or neurotransmitter) that binds to the same receptor.

The term “antagonist,” as used herein, refers to a molecule such as acompound, which diminishes, inhibits, or prevents the action of anothermolecule or the activity of a receptor site. Antagonists include, butare not limited to, competitive antagonists, non-competitiveantagonists, uncompetitive antagonists, partial agonists and inverseagonists.

The term “LPA-dependent”, as used herein, refers to conditions ordisorders that would not occur, or would not occur to the same extent,in the absence of LPA.

The term “LPA-mediated”, as used herein, refers to refers to conditionsor disorders that might occur in the absence of LPA but can occur in thepresence of LPA.

The terms “co-administration” or the like, as used herein, are meant toencompass administration of the selected therapeutic agents to a singlepatient, and are intended to include treatment regimens in which theagents are administered by the same or different route of administrationor at the same or different time.

The terms “effective amount” or “therapeutically effective amount,” asused herein, refer to a sufficient amount of an agent or a compoundbeing administered which will relieve to some extent one or more of thesymptoms of the disease or condition being treated. The result can bereduction and/or alleviation of the signs, symptoms, or causes of adisease, or any other desired alteration of a biological system. Forexample, an “effective amount” for therapeutic uses is the amount of thecomposition comprising a compound as disclosed herein required toprovide a clinically significant decrease in disease symptoms. Anappropriate “effective” amount in any individual case may be determinedusing techniques, such as a dose escalation study.

The terms “enhance” or “enhancing,” as used herein, means to increase orprolong either in potency or duration a desired effect. Thus, in regardto enhancing the effect of therapeutic agents, the term “enhancing”refers to the ability to increase or prolong, either in potency orduration, the effect of other therapeutic agents on a system. An“enhancing-effective amount,” as used herein, refers to an amountadequate to enhance the effect of another therapeutic agent in a desiredsystem.

The terms “kit” and “article of manufacture” are used as synonyms.

A “metabolite” of a compound disclosed herein is a derivative of thatcompound that is formed when the compound is metabolized. The term“active metabolite” refers to a biologically active derivative of acompound that is formed when the compound is metabolized. The term“metabolized,” as used herein, refers to the sum of the processes(including, but not limited to, hydrolysis reactions and reactionscatalyzed by enzymes) by which a particular substance is changed by anorganism. Thus, enzymes may produce specific structural alterations to acompound. For example, cytochrome P450 catalyzes a variety of oxidativeand reductive reactions while uridine diphosphate glucuronyltransferasescatalyze the transfer of an activated glucuronic-acid molecule toaromatic alcohols, aliphatic alcohols, carboxylic acids, amines and freesulphydryl groups. Metabolites of the compounds disclosed herein areoptionally identified either by administration of compounds to a hostand analysis of tissue samples from the host, or by incubation ofcompounds with hepatic cells in vitro and analysis of the resultingcompounds.

The term “pharmaceutical combination” as used herein, means a productthat results from the mixing or combining of more than one activeingredient and includes both fixed and non-fixed combinations of theactive ingredients. The term “fixed combination” means that the activeingredients, e.g. a compound of Formula (I), Formula (II) or Formula(III) and a co-agent, are both administered to a patient simultaneouslyin the form of a single entity or dosage. The term “non-fixedcombination” means that the active ingredients, e.g. a compound ofFormula (I), Formula (II) or Formula (III) and a co-agent, areadministered to a patient as separate entities either simultaneously,concurrently or sequentially with no specific intervening time limits,wherein such administration provides effective levels of the twocompounds in the body of the patient. The latter also applies tococktail therapy, e.g. the administration of three or more activeingredients.

The term “subject” or “patient” encompasses mammals and non-mammals.Examples of mammals include, but are not limited to, any member of theMammalian class: humans, non-human primates such as chimpanzees, andother apes and monkey species; farm animals such as cattle, horses,sheep, goats, swine; domestic animals such as rabbits, dogs, and cats;laboratory animals including rodents, such as rats, mice and guineapigs, and the like. In one embodiment, the mammal is a human.

The terms “treat,” “treating” or “treatment,” as used herein, includealleviating, abating or ameliorating at least one symptom of a diseasedisease or condition, preventing additional symptoms, inhibiting thedisease or condition, e.g., arresting the development of the disease orcondition, relieving the disease or condition, causing regression of thedisease or condition, relieving a condition caused by the disease orcondition, or stopping the symptoms of the disease or condition eitherprophylactically and/or therapeutically.

Routes of Administration

Suitable routes of administration include, but are not limited to, oral,intravenous, rectal, aerosol, parenteral, ophthalmic, pulmonary,transmucosal, transdermal, vaginal, otic, nasal, and topicaladministration. In addition, by way of example only, parenteral deliveryincludes intramuscular, subcutaneous, intravenous, intramedullaryinjections, as well as intrathecal, direct intraventricular,intraperitoneal, intralymphatic, and intranasal injections.

In certain embodiments, a compound as described herein is administeredin a local rather than systemic manner, for example, via injection ofthe compound directly into an organ, often in a depot preparation orsustained release formulation. In specific embodiments, long actingformulations are administered by implantation (for examplesubcutaneously or intramuscularly) or by intramuscular injection.Furthermore, in other embodiments, the drug is delivered in a targeteddrug delivery system, for example, in a liposome coated withorgan-specific antibody. In such embodiments, the liposomes are targetedto and taken up selectively by the organ. In yet other embodiments, thecompound as described herein is provided in the form of a rapid releaseformulation, in the form of an extended release formulation, or in theform of an intermediate release formulation. In yet other embodiments,the compound described herein is administered topically.

Pharmaceutical Compositions/Formulations

In some embodiments, the compounds described herein are formulated intopharmaceutical compositions. Pharmaceutical compositions are formulatedin a conventional manner using one or more pharmaceutically acceptableinactive ingredients that facilitate processing of the active compoundsinto preparations that can be used pharmaceutically. Proper formulationis dependent upon the route of administration chosen. A summary ofpharmaceutical compositions described herein can be found, for example,in Remington: The Science and Practice of Pharmacy, Nineteenth Ed(Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E.,Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical DosageForms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams &Wilkins1999), herein incorporated by reference for such disclosure.

Provided herein are pharmaceutical compositions that include a compoundof Formula (I), Formula (II) or Formula (III) and at least onepharmaceutically acceptable inactive ingredient. In some embodiments,the compounds described herein are administered as pharmaceuticalcompositions in which compounds of Formula (I), Formula (II) and Formula(III) are mixed with other active ingredients, as in combinationtherapy. In other embodiments, the pharmaceutical compositions includeother medicinal or pharmaceutical agents, carriers, adjuvants,preserving, stabilizing, wetting or emulsifying agents, solutionpromoters, salts for regulating the osmotic pressure, and/or buffers. Inyet other embodiments, the pharmaceutical compositions include othertherapeutically valuable substances.

A pharmaceutical composition, as used herein, refers to a mixture of acompound of Formula (I), Formula (II) or Formula (III) with otherchemical components (i.e. pharmaceutically acceptable inactiveingredients), such as carriers, excipients, binders, filling agents,suspending agents, flavoring agents, sweetening agents, disintegratingagents, dispersing agents, surfactants, lubricants, colorants, diluents,solubilizers, moistening agents, plasticizers, stabilizers, penetrationenhancers, wetting agents, anti-foaming agents, antioxidants,preservatives, or one or more combination thereof. The pharmaceuticalcomposition facilitates administration of the compound to an organism.In practicing the methods of treatment or use provided herein,therapeutically effective amounts of compounds described herein areadministered in a pharmaceutical composition to a mammal having adisease, disorder, or condition to be treated. In some embodiments, themammal is a human. A therapeutically effective amount can vary widelydepending on the severity of the disease, the age and relative health ofthe subject, the potency of the compound used and other factors. Thecompounds can be used singly or in combination with one or moretherapeutic agents as components of mixtures.

The pharmaceutical formulations described herein are administered to asubject by appropriate administration routes, including but not limitedto, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular),intranasal, buccal, topical, rectal, or transdermal administrationroutes. The pharmaceutical formulations described herein include, butare not limited to, aqueous liquid dispersions, self-emulsifyingdispersions, solid solutions, liposomal dispersions, aerosols, soliddosage forms, powders, immediate release formulations, controlledrelease formulations, fast melt formulations, tablets, capsules, pills,delayed release formulations, extended release formulations, pulsatilerelease formulations, multiparticulate formulations, and mixed immediateand controlled release formulations.

Pharmaceutical compositions including a compound of Formula (I), Formula(II) or Formula (III) are manufactured in a conventional manner, suchas, by way of example only, by means of conventional mixing, dissolving,granulating, dragee-making, levigating, emulsifying, encapsulating,entrapping or compression processes.

The pharmaceutical compositions will include at least one compound ofFormula (I), Formula (II) or Formula (III) as an active ingredient infree-acid or free-base form, or in a pharmaceutically acceptable saltform. In addition, the methods and pharmaceutical compositions describedherein include the use of N-oxides (if appropriate), crystalline forms,amorphous phases, as well as active metabolites of these compoundshaving the same type of activity. In some embodiments, compoundsdescribed herein exist in unsolvated form or in solvated forms withpharmaceutically acceptable solvents such as water, ethanol, and thelike. The solvated forms of the compounds presented herein are alsoconsidered to be disclosed herein.

In certain embodiments, compositions provided herein include one or morepreservatives to inhibit microbial activity. Suitable preservativesinclude mercury-containing substances such as merfen and thiomersal;stabilized chlorine dioxide; and quaternary ammonium compounds such asbenzalkonium chloride, cetyltrimethylammonium bromide andcetylpyridinium chloride.

In some embodiments, formulations described herein benefit fromantioxidants, metal chelating agents, thiol containing compounds andother general stabilizing agents. Examples of such stabilizing agents,include, but are not limited to: (a) about 0.5% to about 2% w/vglycerol, (b) about 0.1% to about 1% w/v methionine, (c) about 0.1% toabout 2% w/v monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e)about 0.01% to about 2% w/v ascorbic acid, (f) 0.003% to about 0.02% w/vpolysorbate 80, (g) 0.001% to about 0.05% w/v. polysorbate 20, (h)arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrins, (1)pentosan polysulfate and other heparinoids, (m) divalent cations such asmagnesium and zinc; or (n) combinations thereof.

The pharmaceutical compositions described herein, which include acompound of Formula (I), Formula (II) or Formula (III) are formulatedinto any suitable dosage form, including but not limited to, aqueousoral dispersions, liquids, gels, syrups, elixirs, slurries, suspensions,solid oral dosage forms, aerosols, controlled release formulations, fastmelt formulations, effervescent formulations, lyophilized formulations,tablets, powders, pills, dragees, capsules, delayed releaseformulations, extended release formulations, pulsatile releaseformulations, multiparticulate formulations, and mixed immediate releaseand controlled release formulations.

Pharmaceutical preparations for oral use are obtained by mixing one ormore solid excipient with one or more of the compounds described herein,optionally grinding the resulting mixture, and processing the mixture ofgranules, after adding suitable auxiliaries, if desired, to obtaintablets or dragee cores. Suitable excipients include, for example,fillers such as sugars, including lactose, sucrose, mannitol, orsorbitol; cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methylcellulose, microcrystalline cellulose,hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or otherssuch as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. Ifdesired, disintegrating agents are added, such as the cross-linkedcroscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or asalt thereof such as sodium alginate. In some embodiments, dyestuffs orpigments are added to the tablets or dragee coatings for identificationor to characterize different combinations of active compound doses.

Pharmaceutical preparations that are administered orally includepush-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules contain the active ingredients in admixture with filler such aslactose, binders such as starches, and/or lubricants such as talc ormagnesium stearate and, optionally, stabilizers. In soft capsules, theactive compounds are dissolved or suspended in suitable liquids, such asfatty oils, liquid paraffin, or liquid polyethylene glycols. In someembodiments, stabilizers are added.

All formulations for oral administration are in dosages suitable forsuch administration.

In one aspect, solid oral soage forms are prepared by mixing a compoundof Formula (I), Formula (II) or Formula (III) with one or more of thefollowing: antioxidants, flavoring agents, and carrier materials such asbinders, suspending agents, disintegration agents, filling agents,surfactants, solubilizers, stabilizers, lubricants, wetting agents, anddiluents.

In some embodiments, the solid dosage forms disclosed herein are in theform of a tablet, (including a suspension tablet, a fast-melt tablet, abite-disintegration tablet, a rapid-disintegration tablet, aneffervescent tablet, or a caplet), a pill, a powder, a capsule, soliddispersion, solid solution, bioerodible dosage form, controlled releaseformulations, pulsatile release dosage forms, multiparticulate dosageforms, beads, pellets, granules. In other embodiments, thepharmaceutical formulation is in the form of a powder. In still otherembodiments, the pharmaceutical formulation is in the form of a tablet.In other embodiments, pharmaceutical formulations of the compounds ofFormula (I), Formula (II) and Formula (III) are in the form of acapsule.

In some embodiments, solid dosage forms, e.g., tablets, effervescenttablets, and capsules, are prepared by mixing particles of a compound ofFormula (I), Formula (II) or Formula (III) with one or morepharmaceutical excipients to form a bulk blend composition. The bulkblend is readily subdivided into equally effective unit dosage forms,such as tablets, pills, and capsules. In some embodiments, theindividual unit dosages include film coatings. These formulations aremanufactured by conventional formulation techniques.

Conventional formulation techniques include, e.g., one or a combinationof methods: (1) dry mixing, (2) direct compression, (3) milling, (4) dryor non-aqueous granulation, (5) wet granulation, or (6) fusion. Othermethods include, e.g., spray drying, pan coating, melt granulation,granulation, fluidized bed spray drying or coating (e.g., wurstercoating), tangential coating, top spraying, tableting, extruding and thelike.

Suitable carriers for use in the solid dosage forms described hereininclude, but are not limited to, acacia, gelatin, colloidal silicondioxide, calcium glycerophosphate, calcium lactate, maltodextrin,glycerine, magnesium silicate, sodium caseinate, soy lecithin, sodiumchloride, tricalcium phosphate, dipotassium phosphate, sodium stearoyllactylate, carrageenan, monoglyceride, diglyceride, pregelatinizedstarch, hydroxypropylmethylcellulose, hydroxypropylmethylcelluloseacetate stearate, sucrose, microcrystalline cellulose, lactose, mannitoland the like.

Suitable filling agents for use in the solid dosage forms describedherein include, but are not limited to, lactose, calcium carbonate,calcium phosphate, dibasic calcium phosphate, calcium sulfate,microcrystalline cellulose, cellulose powder, dextrose, dextrates,dextran, starches, pregelatinized starch, hydroxypropylmethycellulose(HPMC), hydroxypropylmethycellulose phthalate,hydroxypropylmethylcellulose acetate stearate (HPMCAS), sucrose,xylitol, lactitol, mannitol, sorbitol, sodium chloride, polyethyleneglycol, and the like.

Suitable disintegrants for use in the solid dosage forms describedherein include, but are not limited to, natural starch such as cornstarch or potato starch, a pregelatinized starch, or sodium starchglycolate, a cellulose such as methylcrystalline cellulose,methylcellulose, microcrystalline cellulose, croscarmellose, or across-linked cellulose, such as cross-linked sodiumcarboxymethylcellulose, cross-linked carboxymethylcellulose, orcross-linked croscarmellose, a cross-linked starch such as sodium starchglycolate, a cross-linked polymer such as crospovidone, a cross-linkedpolyvinylpyrrolidone, alginate such as alginic acid or a salt of alginicacid such as sodium alginate, a gum such as agar, guar, locust bean,Karaya, pectin, or tragacanth, sodium starch glycolate, bentonite,sodium lauryl sulfate, sodium lauryl sulfate in combination starch, andthe like.

Binders impart cohesiveness to solid oral dosage form formulations: forpowder filled capsule formulation, they aid in plug formation that canbe filled into soft or hard shell capsules and for tablet formulation,they ensure the tablet remaining intact after compression and helpassure blend uniformity prior to a compression or fill step. Materialssuitable for use as binders in the solid dosage forms described hereininclude, but are not limited to, carboxymethylcellulose,methylcellulose, hydroxypropylmethylcellulose,hydroxypropylmethylcellulose acetate stearate, hydroxyethylcellulose,hydroxypropylcellulose, ethylcellulose, and microcrystalline cellulose,microcrystalline dextrose, amylose, magnesium aluminum silicate,polysaccharide acids, bentonites, gelatin, polyvinylpyrrolidone/vinylacetate copolymer, crospovidone, povidone, starch, pregelatinizedstarch, tragacanth, dextrin, a sugar, such as sucrose, glucose,dextrose, molasses, mannitol, sorbitol, xylitol, lactose, a natural orsynthetic gum such as acacia, tragacanth, ghatti gum, mucilage of isapolhusks, starch, polyvinylpyrrolidone, larch arabogalactan, polyethyleneglycol, waxes, sodium alginate, and the like.

In general, binder levels of 20-70% are used in powder-filled gelatincapsule formulations. Binder usage level in tablet formulations varieswhether direct compression, wet granulation, roller compaction, or usageof other excipients such as fillers which itself can act as moderatebinder. Binder levels of up to 70% in tablet formulations is common.

Suitable lubricants or glidants for use in the solid dosage formsdescribed herein include, but are not limited to, stearic acid, calciumhydroxide, talc, corn starch, sodium stearyl fumerate, alkali-metal andalkaline earth metal salts, such as aluminum, calcium, magnesium, zinc,stearic acid, sodium stearates, magnesium stearate, zinc stearate,waxes, Stearowet®, boric acid, sodium benzoate, sodium acetate, sodiumchloride, leucine, a polyethylene glycol or a methoxypolyethylene glycolsuch as Carbowax™, PEG 4000, PEG 5000, PEG 6000, propylene glycol,sodium oleate, glyceryl behenate, glyceryl palmitostearate, glycerylbenzoate, magnesium or sodium lauryl sulfate, and the like.

Suitable diluents for use in the solid dosage forms described hereininclude, but are not limited to, sugars (including lactose, sucrose, anddextrose), polysaccharides (including dextrates and maltodextrin),polyols (including mannitol, xylitol, and sorbitol), cyclodextrins andthe like.

Suitable wetting agents for use in the solid dosage forms describedherein include, for example, oleic acid, glyceryl monostearate, sorbitanmonooleate, sorbitan monolaurate, triethanolamine oleate,polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitanmonolaurate, quaternary ammonium compounds (e.g., Polyquat 10®), sodiumoleate, sodium lauryl sulfate, magnesium stearate, sodium docusate,triacetin, vitamin E TPGS and the like.

Suitable surfactants for use in the solid dosage forms described hereininclude, for example, sodium lauryl sulfate, sorbitan monooleate,polyoxyethylene sorbitan monooleate, polysorbates, polaxomers, bilesalts, glyceryl monostearate, copolymers of ethylene oxide and propyleneoxide, e.g., Pluronic® (BASF), and the like.

Suitable suspending agents for use in the solid dosage forms describedhere include, but are not limited to, polyvinylpyrrolidone, e.g.,polyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidoneK25, or polyvinylpyrrolidone K30, polyethylene glycol, e.g., thepolyethylene glycol can have a molecular weight of about 300 to about6000, or about 3350 to about 4000, or about 7000 to about 5400, vinylpyrrolidone/vinyl acetate copolymer (S630), sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,polysorbate-80, hydroxyethylcellulose, sodium alginate, gums, such as,e.g., gum tragacanth and gum acacia, guar gum, xanthans, includingxanthan gum, sugars, cellulosics, such as, e.g., sodiumcarboxymethylcellulose, methylcellulose, sodium carboxymethylcellulose,hydroxypropylmethylcellulose, hydroxyethylcellulose, polysorbate-80,sodium alginate, polyethoxylated sorbitan monolaurate, polyethoxylatedsorbitan monolaurate, povidone and the like.

Suitable antioxidants for use in the solid dosage forms described hereininclude, for example, e.g., butylated hydroxytoluene (BHT), sodiumascorbate, and tocopherol.

It should be appreciated that there is considerable overlap betweenadditives used in the solid dosage forms described herein. Thus, theabove-listed additives should be taken as merely exemplary, and notlimiting, of the types of additives that can be included in solid dosageforms of the pharmaceutical compositions described herein. The amountsof such additives can be readily determined by one skilled in the art,according to the particular properties desired.

Compressed tablets are solid dosage forms prepared by compacting thebulk blend of the formulations described above.

In various embodiments, tablets will include one or more flavoringagents.

In other embodiments, the tablets will include a film surrounding thefinal compressed tablet. In some embodiments, the film coating canprovide a delayed release of the compound of Formula (I), Formula (II)or Formula (III) from the formulation. In other embodiments, the filmcoating aids in patient compliance (e.g., Opadry® coatings or sugarcoating). Film coatings including Opadry® typically range from about 1%to about 3% of the tablet weight.

A capsule may be prepared, for example, by placing the bulk blend of theformulation of the compound described above, inside of a capsule. Insome embodiments, the formulations (non-aqueous suspensions andsolutions) are placed in a soft gelatin capsule. In other embodiments,the formulations are placed in standard gelatin capsules or non-gelatincapsules such as capsules comprising HPMC. In other embodiments, theformulation is placed in a sprinkle capsule, wherein the capsule isswallowed whole or the capsule is opened and the contents sprinkled onfood prior to eating.

In various embodiments, the particles of the compound of Formula (I),Formula (II) or Formula (III) and one or more excipients are dry blendedand compressed into a mass, such as a tablet, having a hardnesssufficient to provide a pharmaceutical composition that substantiallydisintegrates within less than about 30 minutes, less than about 35minutes, less than about 40 minutes, less than about 45 minutes, lessthan about 50 minutes, less than about 55 minutes, or less than about 60minutes, after oral administration, thereby releasing the formulationinto the gastrointestinal fluid.

In other embodiments, a powder including a compound of Formula (I),Formula (II) or Formula (III) is formulated to include one or morepharmaceutical excipients and flavors. Such a powder is prepared, forexample, by mixing the the compound of Formula (I), Formula (II) orFormula (III) and optional pharmaceutical excipients to form a bulkblend composition. Additional embodiments also include a suspendingagent and/or a wetting agent. This bulk blend is uniformly subdividedinto unit dosage packaging or multi-dosage packaging units.

In still other embodiments, effervescent powders are also prepared.Effervescent salts have been used to disperse medicines in water fororal administration.

In some embodiments, the pharmaceutical solid oral dosage forms areformulated to provide a controlled release of the compound of Formula(I), Formula (II) or Formula (III). Controlled release refers to therelease of the compound of Formula (I), Formula (II) or Formula (III)from a dosage form in which it is incorporated according to a desiredprofile over an extended period of time. Controlled release profilesinclude, for example, sustained release, prolonged release, pulsatilerelease, and delayed release profiles. In contrast to immediate releasecompositions, controlled release compositions allow delivery of an agentto a subject over an extended period of time according to apredetermined profile. Such release rates can provide therapeuticallyeffective levels of agent for an extended period of time and therebyprovide a longer period of pharmacologic response while minimizing sideeffects as compared to conventional rapid release dosage forms. Suchlonger periods of response provide for many inherent benefits that arenot achieved with the corresponding short acting, immediate releasepreparations.

In some embodiments, the solid dosage forms described herein areformulated as enteric coated delayed release oral dosage forms, i.e., asan oral dosage form of a pharmaceutical composition as described hereinwhich utilizes an enteric coating to affect release in the smallintestine or large intestine. In one aspect, the enteric coated dosageform is a compressed or molded or extruded tablet/mold (coated oruncoated) containing granules, powder, pellets, beads or particles ofthe active ingredient and/or other composition components, which arethemselves coated or uncoated. In one aspect, the enteric coated oraldosage form is in the form of a capsule containing pellets, beads orgranules, which include a compound of Formula (I), Formula (II) orFormula (III), that are coated or uncoated.

Any coatings should be applied to a sufficient thickness such that theentire coating does not dissolve in the gastrointestinal fluids at pHbelow about 5, but does dissolve at pH about 5 and above. Coatings aretypically selected from any of the following:

Shellac—this coating dissolves in media of pH>7; Acrylicpolymers—examples of suitable acrylic polymers include methacrylic acidcopolymers and ammonium methacrylate copolymers. The Eudragit series E,L, S, RL, RS and NE (Rohm Pharma) are available as solubilized inorganic solvent, aqueous dispersion, or dry powders. The Eudragit seriesRL, NE, and RS are insoluble in the gastrointestinal tract but arepermeable and are used primarily for colonic targeting. The Eudragitseries E dissolve in the stomach. The Eudragit series L, L-30D and S areinsoluble in stomach and dissolve in the intestine; Poly Vinyl AcetatePhthalate (PVAP)—PVAP dissolves in pH>5, and it is much less permeableto water vapor and gastric fluids.

Conventional coating techniques such as spray or pan coating areemployed to apply coatings. The coating thickness must be sufficient toensure that the oral dosage form remains intact until the desired siteof topical delivery in the intestinal tract is reached.

In other embodiments, the formulations described herein are deliveredusing a pulsatile dosage form. A pulsatile dosage form is capable ofproviding one or more immediate release pulses at predetermined timepoints after a controlled lag time or at specific sites. Exemplarypulsatile dosage forms and methods of their manufacture are disclosed inU.S. Pat. Nos. 5,011,692, 5,017,381, 5,229,135, 5,840,329 and 5,837,284.In one embodiment, the pulsatile dosage form includes at least twogroups of particles, (i.e. multiparticulate) each containing theformulation described herein. The first group of particles provides asubstantially immediate dose of the compound of Formula (I), Formula(II) or Formula (III) upon ingestion by a mammal. The first group ofparticles can be either uncoated or include a coating and/or sealant. Inone aspect, the second group of particles comprises coated particles.The coating on the second group of particles provides a delay of fromabout 2 hours to about 7 hours following ingestion before release of thesecond dose. Suitable coatings for pharmaceutical compositions aredescribed herein or known in the art.

In some embodiments, pharmaceutical formulations are provided thatinclude particles of a compound of Formula (I), Formula (II) or Formula(III) and at least one dispersing agent or suspending agent for oraladministration to a subject. The formulations may be a powder and/orgranules for suspension, and upon admixture with water, a substantiallyuniform suspension is obtained.

In one aspect, liquid formulation dosage forms for oral administrationare in the form of aqueous suspensions selected from the groupincluding, but not limited to, pharmaceutically acceptable aqueous oraldispersions, emulsions, solutions, elixirs, gels, and syrups. See, e.g.,Singh et al., Encyclopedia of Pharmaceutical Technology, 2nd Ed., pp.754-757 (2002). In addition to the particles of the compound of Formula(I), Formula (II) or Formula (III), the liquid dosage forms includeadditives, such as: (a) disintegrating agents; (b) dispersing agents;(c) wetting agents; (d) at least one preservative, (e) viscosityenhancing agents, (f) at least one sweetening agent, and (g) at leastone flavoring agent. In some embodiments, the aqueous dispersions canfurther include a crystalline inhibitor.

Furthermore, pharmaceutical compositions optionally include one or morepH adjusting agents or buffering agents, including acids such as acetic,boric, citric, lactic, phosphoric and hydrochloric acids; bases such assodium hydroxide, sodium phosphate, sodium borate, sodium citrate,sodium acetate, sodium lactate and tris-hydroxymethylaminomethane; andbuffers such as citrate/dextrose, sodium bicarbonate and ammoniumchloride. Such acids, bases and buffers are included in an amountrequired to maintain pH of the composition in an acceptable range.

Additionally, pharmaceutical compositions optionally include one or moresalts in an amount required to bring osmolality of the composition intoan acceptable range. Such salts include those having sodium, potassiumor ammonium cations and chloride, citrate, ascorbate, borate, phosphate,bicarbonate, sulfate, thiosulfate or bisulfite anions; suitable saltsinclude sodium chloride, potassium chloride, sodium thiosulfate, sodiumbisulfite and ammonium sulfate.

Other pharmaceutical compositions optionally include one or morepreservatives to inhibit microbial activity. Suitable preservativesinclude mercury-containing substances such as merfen and thiomersal;stabilized chlorine dioxide; and quaternary ammonium compounds such asbenzalkonium chloride, cetyltrimethylammonium bromide andcetylpyridinium chloride.

In one embodiment, the aqueous suspensions and dispersions describedherein remain in a homogenous state, as defined in The USP Pharmacists'Pharmacopeia (2005 edition, chapter 905), for at least 4 hours. In oneembodiment, an aqueous suspension is re-suspended into a homogenoussuspension by physical agitation lasting less than 1 minute. In stillanother embodiment, no agitation is necessary to maintain a homogeneousaqueous dispersion.

Examples of disintegrating agents for use in the aqueous suspensions anddispersions include, but are not limited to, a starch, e.g., a naturalstarch such as corn starch or potato starch, a pregelatinized starch, orsodium starch glycolate; a cellulose such as methylcrystallinecellulose, methylcellulose, croscarmellose, or a cross-linked cellulose,such as cross-linked sodium carboxymethylcellulose, cross-linkedcarboxymethylcellulose, or cross-linked croscarmellose; a cross-linkedstarch such as sodium starch glycolate; a cross-linked polymer such ascrospovidone; a cross-linked polyvinylpyrrolidone; alginate such asalginic acid or a salt of alginic acid such as sodium alginate; a gumsuch as agar, guar, locust bean, Karaya, pectin, or tragacanth; sodiumstarch glycolate; bentonite; a natural sponge; a surfactant; a resinsuch as a cation-exchange resin; citrus pulp; sodium lauryl sulfate;sodium lauryl sulfate in combination starch; and the like.

In some embodiments, the dispersing agents suitable for the aqueoussuspensions and dispersions described herein include, for example,hydrophilic polymers, electrolytes, Tween® 60 or 80, PEG,polyvinylpyrrolidone, and the carbohydrate-based dispersing agents suchas, for example, hydroxypropylcellulose and hydroxypropyl celluloseethers, hydroxypropyl methylcellulose and hydroxypropyl methylcelluloseethers, carboxymethylcellulose sodium, methylcellulose,hydroxyethylcellulose, hydroxypropylmethyl-cellulose phthalate,hydroxypropylmethyl-cellulose acetate stearate, noncrystallinecellulose, magnesium aluminum silicate, triethanolamine, polyvinylalcohol (PVA), polyvinylpyrrolidone/vinyl acetate copolymer,4-(1,1,3,3-tetramethylbutyl)-phenol polymer with ethylene oxide andformaldehyde (also known as tyloxapol), poloxamers; and poloxamines. Inother embodiments, the dispersing agent is selected from a group notcomprising one of the following agents: hydrophilic polymers;electrolytes; Tween® 60 or 80; PEG; polyvinylpyrrolidone (PVP);hydroxypropylcellulose and hydroxypropyl cellulose ethers; hydroxypropylmethylcellulose and hydroxypropyl methylcellulose ethers;carboxymethylcellulose sodium; methylcellulose; hydroxyethylcellulose;hydroxypropylmethyl-cellulose phthalate; hydroxypropylmethyl-celluloseacetate stearate; non-crystalline cellulose; magnesium aluminumsilicate; triethanolamine; polyvinyl alcohol (PVA);4-(1,1,3,3-tetramethylbutyl)-phenol polymer with ethylene oxide andformaldehyde; poloxamers; or poloxamines.

Wetting agents suitable for the aqueous suspensions and dispersionsdescribed herein include, but are not limited to, cetyl alcohol,glycerol monostearate, polyoxyethylene sorbitan fatty acid esters (e.g.,the commercially available Tweens® such as e.g., Tween 20® and Tween80®, and polyethylene glycols, oleic acid, glyceryl monostearate,sorbitan monooleate, sorbitan monolaurate, triethanolamine oleate,polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitanmonolaurate, sodium oleate, sodium lauryl sulfate, sodium docusate,triacetin, vitamin E TPGS, sodium taurocholate, simethicone,phosphotidylcholine and the like.

Suitable preservatives for the aqueous suspensions or dispersionsdescribed herein include, for example, potassium sorbate, parabens(e.g., methylparaben and propylparaben), benzoic acid and its salts,other esters of parahydroxybenzoic acid such as butylparaben, alcoholssuch as ethyl alcohol or benzyl alcohol, phenolic compounds such asphenol, or quaternary compounds such as benzalkonium chloride.Preservatives, as used herein, are incorporated into the dosage form ata concentration sufficient to inhibit microbial growth.

Suitable viscosity enhancing agents for the aqueous suspensions ordispersions described herein include, but are not limited to, methylcellulose, xanthan gum, carboxymethyl cellulose, hydroxypropylcellulose, hydroxypropylmethyl cellulose, Plasdon® S-630, carbomer,polyvinyl alcohol, alginates, acacia, chitosans and combinationsthereof. The concentration of the viscosity enhancing agent will dependupon the agent selected and the viscosity desired.

Examples of sweetening agents suitable for the aqueous suspensions ordispersions described herein include, for example, acacia syrup,acesulfame K, alitame, aspartame, chocolate, cinnamon, citrus, cocoa,cyclamate, dextrose, fructose, ginger, glycyrrhetinate, glycyrrhiza(licorice) syrup, monoammonium glyrrhizinate (MagnaSweet®), maltol,mannitol, menthol, neohesperidine DC, neotame, Prosweet® Powder,saccharin, sorbitol, stevia, sucralose, sucrose, sodium saccharin,saccharin, aspartame, acesulfame potassium, mannitol, sucralose,tagatose, thaumatin, vanilla, xylitol, or any combination thereof.

In some embodiments, the liquid formulations also include inert diluentscommonly used in the art, such as water or other solvents, solubilizingagents, and emulsifiers. Exemplary emulsifiers are ethyl alcohol,isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol,benzyl benzoate, propyleneglycol, 1,3-butyleneglycol, dimethylformamide,sodium lauryl sulfate, sodium doccusate, cholesterol, cholesterolesters, taurocholic acid, phosphotidylcholine, oils, such as cottonseedoil, groundnut oil, corn germ oil, olive oil, castor oil, and sesameoil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols, fattyacid esters of sorbitan, or mixtures of these substances, and the like.

Representative intranasal formulations are described in, for example,U.S. Pat. Nos. 4,476,116, 5,116,817 and 6,391,452. Formulations thatinclude a compound of Formula (I), Formula (II) or Formula (III) areprepared as solutions in saline, employing benzyl alcohol or othersuitable preservatives, fluorocarbons, and/or other solubilizing ordispersing agents known in the art. See, for example, Ansel, H. C. etal., Pharmaceutical Dosage Forms and Drug Delivery Systems, Sixth Ed.(1995). Preferably these compositions and formulations are prepared withsuitable nontoxic pharmaceutically acceptable ingredients. Theseingredients are known to those skilled in the preparation of nasaldosage forms and some of these can be found in REMINGTON: THE SCIENCEAND PRACTICE OF PHARMACY, 21st edition, 2005. The choice of suitablecarriers is dependent upon the exact nature of the nasal dosage formdesired, e.g., solutions, suspensions, ointments, or gels. Nasal dosageforms generally contain large amounts of water in addition to the activeingredient. Minor amounts of other ingredients such as pH adjusters,emulsifiers or dispersing agents, preservatives, surfactants, gellingagents, or buffering and other stabilizing and solubilizing agents areoptionally present. Preferably, the nasal dosage form should be isotonicwith nasal secretions.

For administration by inhalation, a compound of Formula (I), Formula(II) or Formula (III) is formulated for use as an aerosol, a mist or apowder. Pharmaceutical compositions described herein are convenientlydelivered in the form of an aerosol spray presentation from pressurizedpacks or a nebuliser, with the use of a suitable propellant, e.g.,dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of a pressurized aerosol, the dosage unit may be determined byproviding a valve to deliver a metered amount. Capsules and cartridgesof, such as, by way of example only, gelatin for use in an inhaler orinsufflator may be formulated containing a powder mix of the compounddescribed herein and a suitable powder base such as lactose or starch.

Buccal formulations that include a compound of Formula (I), Formula (II)or Formula (III) are administered using a variety of formulations knownin the art. For example, such formulations include, but are not limitedto, U.S. Pat. Nos. 4,229,447, 4,596,795, 4,755,386, and 5,739,136. Inaddition, the buccal dosage forms described herein can further include abioerodible (hydrolysable) polymeric carrier that also serves to adherethe dosage form to the buccal mucosa. For buccal or sublingualadministration, the compositions may take the form of tablets, lozenges,or gels formulated in a conventional manner.

In some embodiments, compounds of Formula (I), Formula (II) and Formula(III) are prepared as transdermal dosage forms. In one embodiments, thetransdermal formulations described herein include at least threecomponents: (1) a formulation of a compound of Formula (I), Formula (II)or Formula (III); (2) a penetration enhancer; and (3) an aqueousadjuvant. In some embodiments the transdermal formulations includeadditional components such as, but not limited to, gelling agents,creams and ointment bases, and the like. In some embodiments, thetransdermal formulation further include a woven or non-woven backingmaterial to enhance absorption and prevent the removal of thetransdermal formulation from the skin. In other embodiments, thetransdermal formulations described herein can maintain a saturated orsupersaturated state to promote diffusion into the skin.

In one aspect, formulations suitable for transdermal administration ofcompounds described herein employ transdermal delivery devices andtransdermal delivery patches and can be lipophilic emulsions orbuffered, aqueous solutions, dissolved and/or dispersed in a polymer oran adhesive. In one aspect, such patches are constructed for continuous,pulsatile, or on demand delivery of pharmaceutical agents. Stillfurther, transdermal delivery of the compounds described herein can beaccomplished by means of iontophoretic patches and the like. In oneaspect, transdermal patches provide controlled delivery of the compoundof Formula (I), Formula (II) or Formula (III). In one aspect,transdermal devices are in the form of a bandage comprising a backingmember, a reservoir containing the compound optionally with carriers,optionally a rate controlling barrier to deliver the compound to theskin of the host at a controlled and predetermined rate over a prolongedperiod of time, and means to secure the device to the skin.

In one aspect, a compound of Formula (I), Formula (II) or Formula (III)is formulated into a pharmaceutical composition suitable forintramuscular, subcutaneous, or intravenous injection. In one aspect,formulations suitable for intramuscular, subcutaneous, or intravenousinjection include physiologically acceptable sterile aqueous ornon-aqueous solutions, dispersions, suspensions or emulsions, andsterile powders for reconstitution into sterile injectable solutions ordispersions. Examples of suitable aqueous and non-aqueous carriers,diluents, solvents, or vehicles include water, ethanol, polyols(propyleneglycol, polyethylene-glycol, glycerol, cremophor and thelike), suitable mixtures thereof, vegetable oils (such as olive oil) andinjectable organic esters such as ethyl oleate. Proper fluidity can bemaintained, for example, by the use of a coating such as lecithin, bythe maintenance of the required particle size in the case ofdispersions, and by the use of surfactants. In some embodiments,formulations suitable for subcutaneous injection also contain additivessuch as preserving, wetting, emulsifying, and dispensing agents.Prevention of the growth of microorganisms can be ensured by variousantibacterial and antifungal agents, such as parabens, chlorobutanol,phenol, sorbic acid, and the like. In some cases it is desirable toinclude isotonic agents, such as sugars, sodium chloride, and the like.Prolonged absorption of the injectable pharmaceutical form can bebrought about by the use of agents delaying absorption, such as aluminummonostearate and gelatin.

For intravenous injections, compounds described herein are formulated inaqueous solutions, preferably in physiologically compatible buffers suchas Hank's solution, Ringer's solution, or physiological saline buffer.For transmucosal administration, penetrants appropriate to the barrierto be permeated are used in the formulation. Such penetrants aregenerally known in the art. For other parenteral injections, appropriateformulations include aqueous or nonaqueous solutions, preferably withphysiologically compatible buffers or excipients. Such excipients areknown.

Parenteral injections may involve bolus injection or continuousinfusion. Formulations for injection may be presented in unit dosageform, e.g., in ampoules or in multi-dose containers, with an addedpreservative. The pharmaceutical composition described herein may be ina form suitable for parenteral injection as a sterile suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilizing and/or dispersingagents. In one aspect, the active ingredient is in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use.

In certain embodiments, delivery systems for pharmaceutical compoundsmay be employed, such as, for example, liposomes and emulsions. Incertain embodiments, compositions provided herein can also include anmucoadhesive polymer, selected from among, for example,carboxymethylcellulose, carbomer (acrylic acid polymer),poly(methylmethacrylate), polyacrylamide, polycarbophil, acrylicacid/butyl acrylate copolymer, sodium alginate and dextran.

In some embodiments, the compounds described herein may be administeredtopically and can be formulated into a variety of topicallyadministrable compositions, such as solutions, suspensions, lotions,gels, pastes, medicated sticks, balms, creams or ointments. Suchpharmaceutical compounds can contain solubilizers, stabilizers, tonicityenhancing agents, buffers and preservatives.

In some embodiments, the compounds of Formula (I), Formula (II) andFormula (III) are formulated in rectal compositions such as enemas,rectal gels, rectal foams, rectal aerosols, suppositories, jellysuppositories, or retention enemas, containing conventional suppositorybases such as cocoa butter or other glycerides, as well as syntheticpolymers such as polyvinylpyrrolidone, PEG, and the like. In suppositoryforms of the compositions, a low-melting wax such as, but not limitedto, a mixture of fatty acid glycerides, optionally in combination withcocoa butter is first melted.

Methods of Dosing and Treatment Regimens

In one embodiment, the compounds of Formula (I), Formula (II) andFormula (III) are used in the preparation of medicaments for thetreatment of LPA-dependent or LPA-mediated diseases or conditions. Inaddition, a method for treating any of the diseases or conditionsdescribed herein in a subject in need of such treatment, involvesadministration of pharmaceutical compositions that include at least onecompound of Formula (I), Formula (II) or Formula (III) or apharmaceutically acceptable salt, pharmaceutically active metabolite,pharmaceutically acceptable prodrug, or pharmaceutically acceptablesolvate thereof, in therapeutically effective amounts to said subject.

In certain embodiments, the compositions containing the compound(s)described herein are administered for prophylactic and/or therapeutictreatments. In certain therapeutic applications, the compositions areadministered to a patient already suffering from a disease or condition,in an amount sufficient to cure or at least partially arrest at leastone of the symptoms of the disease or condition. Amounts effective forthis use depend on the severity and course of the disease or condition,previous therapy, the patient's health status, weight, and response tothe drugs, and the judgment of the treating physician. Therapeuticallyeffective amounts are optionally determined by methods including, butnot limited to, a dose escalation clinical trial.

In prophylactic applications, compositions containing the compoundsdescribed herein are administered to a patient susceptible to orotherwise at risk of a particular disease, disorder or condition. Suchan amount is defined to be a “prophylactically effective amount ordose.” In this use, the precise amounts also depend on the patient'sstate of health, weight, and the like. When used in a patient, effectiveamounts for this use will depend on the severity and course of thedisease, disorder or condition, previous therapy, the patient's healthstatus and response to the drugs, and the judgment of the treatingphysician. In one aspect, prophylactic treatments include administeringto a mammal, who previously experienced at least one symptom of thedisease being treated and is currently in remission, a pharmaceuticalcomposition comprising a compound of Formula (I), Formula (II) orFormula (III) in order to prevent a return of the symptoms of thedisease or condition.

In certain embodiments wherein the patient's condition does not improve,upon the doctor's discretion the administration of the compounds areadministered chronically, that is, for an extended period of time,including throughout the duration of the patient's life in order toameliorate or otherwise control or limit the symptoms of the patient'sdisease or condition.

In certain embodiments wherein a patient's status does improve, the doseof drug being administered may be temporarily reduced or temporarilysuspended for a certain length of time (i.e., a “drug holiday”). Inspecific embodiments, the length of the drug holiday is between 2 daysand 1 year, including by way of example only, 2 days, 3 days, 4 days, 5days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, ormore than 28 days. The dose reduction during a drug holiday is, by wayof example only, by 10%-100%, including by way of example only 10%, 15%,20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, and 100%.

In certain embodiments in which the patient is presented with asituation in which the activity of LPA needs to be enhanced, forexample, to assist with wound healing, the dose of drug beingadministered may be temporarily reduced or temporarily suspended for acertain length of time (i.e., a “drug diversion”). In specificembodiments, the length of the drug diversion is between 2 days and 1year, including by way of example only, 2 days, 3 days, 4 days, 5 days,6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, or morethan 28 days. The dose reduction during a drug diversion is, by way ofexample only, by 10%-100%, including by way of example only 10%, 15%,20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, and 100%. Once the situation requiring enhanced activity ofLPA is alleviated, the normal dosing schedule is optionally reinstated.

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, in specificembodiments, the dosage or the frequency of administration, or both, isreduced, as a function of the symptoms, to a level at which the improveddisease, disorder or condition is retained. In certain embodiments,however, the patient requires intermittent treatment on a long-termbasis upon any recurrence of symptoms.

The amount of a given agent that corresponds to such an amount variesdepending upon factors such as the particular compound, diseasecondition and its severity, the identity (e.g., weight, sex) of thesubject or host in need of treatment, but can nevertheless be determinedaccording to the particular circumstances surrounding the case,including, e.g., the specific agent being administered, the route ofadministration, the condition being treated, and the subject or hostbeing treated. In general, however, doses employed for adult humantreatment are typically in the range of 0.01 mg-5000 mg per day. In oneaspect, doses employed for adult human treatment are from about lmg toabout 1000 mg per day. In one embodiment, the desired dose isconveniently presented in a single dose or in divided doses administeredsimultaneously (or over a short period of time) or at appropriateintervals, for example as two, three, four or more sub-doses per day.

In one embodiment, the daily dosages appropriate for the compound ofFormula (I), Formula (II) or Formula (III) described herein are fromabout 0.01 to about 10 mg/kg per body weight. In specific embodiments,an indicated daily dosage in a large mammal, including, but not limitedto, humans, is in the range from about 0.5 mg to about 1000 mg,conveniently administered in divided doses, including, but not limitedto, up to four times a day. In one embodiment, the daily dosage isadministered in extended release form. In certain embodiments, suitableunit dosage forms for oral administration comprise from about 1 to 500mg active ingredient. In other embodiments, the daily dosage or theamount of active in the dosage form are lower or higher than the rangesindicated herein, based on a number of variables in regard to anindividual treatment regime. In various embodiments, the daily and unitdosages are altered depending on a number of variables including, butnot limited to, the activity of the compound used, the disease orcondition to be treated, the mode of administration, the requirements ofthe individual subject, the severity of the disease or condition beingtreated, and the judgment of the practitioner.

Toxicity and therapeutic efficacy of such therapeutic regimens aredetermined by standard pharmaceutical procedures in cell cultures orexperimental animals, including, but not limited to, the determinationof the LD₅₀ and the ED₅₀. The dose ratio between the toxic andtherapeutic effects is the therapeutic index and it is expressed as theratio between LD₅₀ and ED₅₀. In certain embodiments, the data obtainedfrom cell culture assays and animal studies are used in formulating thetherapeutically effective daily dosage range and/or the therapeuticallyeffective unit dosage amount for use in mammals, including humans. Insome embodiments, the daily dosage amount of the compounds describedherein lies within a range of circulating concentrations that includethe ED₅₀ with minimal toxicity. In certain embodiments, the daily dosagerange and/or the unit dosage amount varies within this range dependingupon the dosage form employed and the route of administration utilized.

Patient Selection

In any of the aforementioned aspects involving the prevention ortreatment of LPA-mediated diseases or conditions are further embodimentscomprising identifying patients by screening for LPA receptor gene SNPs.A SNP located in the promoter region of LPA₁ showed significantassociation with knee osteoarthritis in two independent populations(Mototani et al. Hum. Mol. Genetics, vol. 17, no. 12, 2008). Patientscan be further selected based on increased LPA receptor expression inthe tissue of interest. For example, chronic lymphocytic leukemia (CLL)is characterized by the accumulation of CD19+/CD5+ B-lymphocytes in theperipheral blood, bone marrow and lymphoid organs which occurs as aresult of a block in B-lymphocyte apoptosis. LPA can protect some CLLcells from apoptosis and the cells that are protected by LPA have highlevels of LPA₁ mRNA. In some embodiments, CLL patients are selectedbased on the expression of the LPA1R. LPA receptor expression aredetermined by methods including, but not limited to, northern blotting,western blotting, quantitative PCR (qPCR), flow cytometry,autoradiography (using a small molecule radioligand or PET ligand). Insome embodiments, patients are selected based on the concentration ofserum or tissue LPA measured by mass spectrometry. LPA concentrationsare high in ovarian cancer ascites and in some breast cancer effusions.In some embodiments, patients are selected based on a combination of theabove markers (increased LPA concentrations and increased LPA receptorexpression).

Combination Treatments

In certain instances, it is appropriate to administer at least onecompound of Formula (I), Formula (II) or Formula (III) in combinationwith another therapeutic agent. By way of example only, if one of theside effects experienced by a patient upon receiving one of thecompounds herein is inflammation, then it may be appropriate toadminister an anti-inflammatory agent in combination with the initialtherapeutic agent. Or in another example, a patient is presented with asituation in which antagonism of LPA receptors provides potential harm,for example, if the patient is wounded, antagonism of LPA receptors maylead to a delay in wound healing. In such an event, in certainembodiments, the patient benefits by administration of a localwound-healing agent (at the site of the wound) in combination with theco-existing administration of a compound of Formula (I), Formula (II) orFormula (III).

Or, in one embodiment, the therapeutic effectiveness of one of thecompounds described herein is enhanced by administration of an adjuvant(i.e., by itself the adjuvant may have minimal therapeutic benefit, butin combination with another therapeutic agent, the overall therapeuticbenefit to the patient is enhanced). Or, in some embodiments, thebenefit experienced by a patient is increased by administering one ofthe compounds described herein with another therapeutic agent (whichalso includes a therapeutic regimen) that also has therapeutic benefit.

In one specific embodiment, a compound of Formula (I), Formula (II) orFormula (III) is co-administered with a second therapeutic agent,wherein the compound of Formula (I), Formula (II) or Formula (III) andthe second therapeutic agent modulate different aspects of the disease,disorder or condition being treated, thereby providing a greater overallbenefit than administration of either therapeutic agent alone.

In any case, regardless of the disease, disorder or condition beingtreated, the overall benefit experienced by the patient may simply beadditive of the two therapeutic agents or the patient may experience asynergistic benefit.

In certain embodiments, different therapeutically-effective dosages ofthe compounds disclosed herein will be utilized in formulatingpharmaceutical composition and/or in treatment regimens when thecompounds disclosed herein are administered in combination with one ormore additional agent, such as an additional therapeutically effectivedrug, an adjuvant or the like. Therapeutically-effective dosages ofdrugs and other agents for use in combination treatment regimens can bedetermined by means similar to those set forth hereinabove for theactives themselves. Furthermore, the methods of prevention/treatmentdescribed herein encompasses the use of metronomic dosing, i.e.,providing more frequent, lower doses in order to minimize toxic sideeffects. In some embodiments, a combination treatment regimenencompasses treatment regimens in which administration of a compound ofFormula (I), Formula (II) or Formula (III) is initiated prior to,during, or after treatment with a second agent described herein, andcontinues until any time during treatment with the second agent or aftertermination of treatment with the second agent. It also includestreatments in which a compound of Formula (I), Formula (II) or Formula(III) and the second agent being used in combination are administeredsimultaneously or at different times and/or at decreasing or increasingintervals during the treatment period. Combination treatment furtherincludes periodic treatments that start and stop at various times toassist with the clinical management of the patient.

Compositions and methods for combination therapy are provided herein. Inaccordance with one aspect, the pharmaceutical compositions disclosedherein are used to treat LPA-dependent or LPA-mediated conditions.

It is understood that the dosage regimen to treat, prevent, orameliorate the condition(s) for which relief is sought, is modified inaccordance with a variety of factors. These factors include the disease,disorder or condition from which the subject suffers, as well as theage, weight, sex, diet, and medical condition of the subject. Thus, insome instances, the dosage regimen actually employed varies and, in someembodiments, deviates from the dosage regimens set forth herein.

For combination therapies described herein, dosages of theco-administered compounds vary depending on the type of co-drugemployed, on the specific drug employed, on the disease or conditionbeing treated and so forth. In additional embodiments, whenco-administered with one or more other therapeutic agents, the compoundprovided herein is administered either simultaneously with the one ormore other therapeutic agents, or sequentially.

In combination therapies, the multiple therapeutic agents (one of whichis one of the compounds described herein) are administered in any orderor even simultaneously. If administration is simultaneous, the multipletherapeutic agents are, by way of example only, provided in a single,unified form, or in multiple forms (e.g., as a single pill or as twoseparate pills). In one embodiment, one of the therapeutic agents isgiven in multiple doses, and in another, two (or more if present) aregiven as multiple doses. In some embodiments of non-simultaneousadministration, the timing between the multiple doses vary from morethan zero weeks to less than four weeks. In addition, the combinationmethods, compositions and formulations are not to be limited to the useof only two agents; the use of multiple therapeutic combinations is alsoenvisioned.

The compounds of Formula (I), Formula (II) and Formula (III), andcombination therapies, are administered before, during or after theoccurrence of a disease or condition, and the timing of administeringthe composition containing a compound varies. Thus, in one embodiment,the compounds described herein are used as a prophylactic and areadministered continuously to subjects with a propensity to developconditions or diseases in order to prevent the occurrence of the diseaseor condition. In another embodiment, the compounds and compositions areadministered to a subject during or as soon as possible after the onsetof the symptoms. In specific embodiments, a compound described herein isadministered as soon as is practicable after the onset of a disease orcondition is detected or suspected, and for a length of time necessaryfor the treatment of the disease. In some embodiments, the lengthrequired for treatment varies, and the treatment length is adjusted tosuit the specific needs of each subject. For example, in specificembodiments, a compound described herein or a formulation containing thecompound is administered for at least 2 weeks, about 1 month to about 5years.

By way of example, therapies which combine a compound of Formula (I),Formula (II) or Formula (III) with inhibitors of LPA synthesis or LPAreceptor antagonists, either acting at the same or other points in theLPA synthesis or signalling pathway, are encompassed herein for treatingLPA-dependent or LPA-mediated diseases or conditions.

Exemplary Agents for Use in Combination Therapy

In another embodiment described herein, methods for treatment ofLPA-dependent or LPA-mediated conditions or diseases, such asproliferative disorders, including cancer, comprises administration to amammal a compound of Formula (I), Formula (II) or Formula (III) incombination with at least one additional agent selected, by way ofexample only, alemtuzumab, arsenic trioxide, asparaginase (pegylated ornon-), bevacizumab, cetuximab, platinum-based compounds such ascisplatin, cladribine, daunorubicin/doxorubicin/idarubicin, irinotecan,fludarabine, 5-fluorouracil, gemtuzumab, methotrexate, Paclitaxel™,taxol, temozolomide, thioguanine, or classes of drugs including hormones(an antiestrogen, an antiandrogen, or gonadotropin releasing hormoneanalogues, interferons such as alpha interferon, nitrogen mustards suchas busulfan or melphalan or mechlorethamine, retinoids such astretinoin, topoisomerase inhibitors such as irinotecan or topotecan,tyrosine kinase inhibitors such as gefinitinib or imatinib, or agents totreat signs or symptoms induced by such therapy including allopurinol,filgrastim, granisetron/ondansetron/palonosetron, dronabinol.

In one aspect, the compound of Formula (I), Formula (II) or Formula(III) is administered or formulated in combination with one or moreanti-cancer agents. In some embodiments, one or more of the anti-canceragents are proapoptotic agents. Examples of anti-cancer agents include,but are not limited to, any of the following: gossypol, genasense,polyphenol E, Chlorofusin, all trans-retinoic acid (ATRA), bryostatin,tumor necrosis factor-related apoptosis-inducing ligand (TRAIL),5-aza-2′-deoxycytidine, all trans retinoic acid, doxorubicin,vincristine, etoposide, gemcitabine, imatinib, geldanamycin,17-N-Allylamino-17-Demethoxygeldanamycin (17-AAG), flavopiridol,LY294002, bortezomib, trastuzumab, BAY 11-7082, PKC412, or PD184352,Taxol™ (paclitaxel), and analogs of Taxol™, such as Taxotere™. Compoundsthat have the basic taxane skeleton as a common structure feature, havealso been shown to have the ability to arrest cells in the G2-M phasesdue to stabilized microtubules and may be useful for treating cancer incombination with the compounds described herein.

Further examples of anti-cancer agents for use in combination with acompound of Formula (I), Formula (II) or Formula (III) includeinhibitors of mitogen-activated protein kinase signaling, e.g., U0126,PD98059, PD184352, PD0325901, ARRY-142886, SB239063, SP600125, BAY43-9006, wortmannin, or LY294002; Syk inhibitors; mTOR inhibitors; andantibodies (e.g., rituxan).

Other anti-cancer agents for use in combination with a compound ofFormula (I), Formula (II) or Formula (III) include one or more of thefollowing: Adriamycin, Dactinomycin, Bleomycin, Vinblastine, Cisplatin,acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin;aldesleukin; altretamine; ambomycin; ametantrone acetate;aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase;asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa;bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin;bleomycin sulfate; brequinar sodium; bropirimine; busulfan;cactinomycin; calusterone; caracemide; carbetimer; carboplatin;carmustine; carubicin hydrochloride; carzelesin; cedefingol;chlorambucil; cirolemycin; cladribine; crisnatol mesylate;cyclophosphamide; cytarabine; dacarbazine; daunorubicin hydrochloride;decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate;diaziquone; doxorubicin; doxorubicin hydrochloride; droloxifene;droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate;eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate;epipropidine; epirubicin hydrochloride; erbulozole; esorubicinhydrochloride; estramustine; estramustine phosphate sodium; etanidazole;etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride;fazarabine; fenretinide; floxuridine; fludarabine phosphate;fluorouracil; flurocitabine; fosquidone; fostriecin sodium; gemcitabine;gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride;ifosfamide; iimofosine; interleukin I1 (including recombinantinterleukin II, or rlL2), interferon alfa-2a; interferon alfa-2b;interferon alfa-n1; interferon alfa-n3; interferon beta-1 a; interferongamma-1 b; iproplatin; irinotecan hydrochloride; lanreotide acetate;letrozole; leuprolide acetate; liarozole hydrochloride; lometrexolsodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine;mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate;melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium;metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin;mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride;mycophenolic acid; nocodazoie; nogalamycin; ormaplatin; oxisuran;pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride;semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantronehydrochloride; temoporfin; teniposide; teroxirone; testolactone;thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifenecitrate; trestolone acetate; triciribine phosphate; trimetrexate;trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracilmustard; uredepa; vapreotide; verteporfin; vinblastine sulfate;vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate;vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate;vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin;zinostatin; zorubicin hydrochloride.

Yet other anticancer agents for use in combination with a compound ofFormula (I), Formula (II) or Formula (III) include alkylating agents,antimetabolites, natural products, or hormones, e.g., nitrogen mustards(e.g., mechloroethamine, cyclophosphamide, chlorambucil, etc.), alkylsulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine, lomusitne,ete.), or triazenes (decarbazine, etc.). Examples of antimetabolitesinclude but are not limited to folic acid analog (e.g., methotrexate),or pyrimidine analogs (e.g., Cytarabine), purine analogs (e.g.,mercaptopurine, thioguanine, pentostatin).

Examples of natural products for use in combination with a compound ofFormula (I), Formula (II) or Formula (III) include but are not limitedto vinca alkaloids (e.g., vinblastin, vincristine), epipodophyllotoxins(e.g., etoposide), antibiotics (e.g., daunorubicin, doxorubicin,bleomycin), enzymes (e.g., L-asparaginase), or biological responsemodifiers (e.g., interferon alpha).

Examples of alkylating agents for use in combination with a compound ofFormula (I), Formula (II) or Formula (III) include, but are not limitedto, nitrogen mustards (e.g., mechloroethamine, cyclophosphamide,chlorambucil, meiphalan, etc.), ethylenimine and methylmelamines (e.g.,hexamethlymelamine, thiotepa), alkyl sulfonates (e.g., busulfan),nitrosoureas (e.g., carmustine, lomusitne, semustine, streptozocin,etc.), or triazenes (decarbazine, ete.). Examples of antimetabolitesinclude, but are not limited to folic acid analog (e.g., methotrexate),or pyrimidine analogs (e.g., fluorouracil, floxouridine, Cytarabine),purine analogs (e.g., mercaptopurine, thioguanine, pentostatin.

Examples of hormones and antagonists for use in combination with acompound of Formula (I), Formula (II) or Formula (III) include, but arenot limited to, adrenocorticosteroids (e.g., prednisone), progestins(e.g., hydroxyprogesterone caproate, megestrol acetate,medroxyprogesterone acetate), estrogens (e.g., diethlystilbestrol,ethinyl estradiol), antiestrogen (e.g., tamoxifen), androgens (e.g.,testosterone propionate, fluoxymesterone), antiandrogen (e.g.,flutamide), gonadotropin releasing hormone analog (e.g., leuprolide).Other agents that can be used in the methods and compositions describedherein for the treatment or prevention of cancer include platinumcoordination complexes (e.g., cisplatin, carboblatin), anthracenedione(e.g., mitoxantrone), substituted urea (e.g., hydroxyurea), methylhydrazine derivative (e.g., procarbazine), adrenocortical suppressant(e.g., mitotane, aminoglutethimide).

Examples of anti-cancer agents which act by arresting cells in the G2-Mphases due to stabilized microtubules include without limitation thefollowing marketed drugs and drugs in development: Erbulozole,Dolastatin 10, Mivobulin isethionate, Vincristine, NSC-639829,Discodermolide, ABT-751, Altorhyrtins (such as Altorhyrtin A andAltorhyrtin C), Spongistatins (such as Spongistatin 1, Spongistatin 2,Spongistatin 3, Spongistatin 4, Spongistatin 5, Spongistatin 6,Spongistatin 7, Spongistatin 8, and Spongistatin 9), Cemadotinhydrochloride, Epothilones (such as Epothilone A, Epothilone B,Epothilone C, Epothilone D, Epothilone E, Epothilone F, Epothilone BN-oxide, Epothilone A N-oxide, 16-aza-epothilone B, 21-aminoepothiloneB, 21-hydroxyepothilone D, 26-fluoroepothilone, Auristatin PE,Soblidotin, Vincristine sulfate, Cryptophycin 52, Vitilevuamide,Tubulysin A, Canadensol, Centaureidin, Oncocidin Al Fijianolide B,Laulimalide, Narcosine, Nascapine, Hemiasterlin, Vanadoceneacetylacetonate, Indanocine Eleutherobins (such asDesmethyleleutherobin, Desaetyleleutherobin, lsoeleutherobin A, andZ-Eleutherobin), Caribaeoside, Caribaeolin, Halichondrin B, DiazonamideA, Taccalonolide A, Diozostatin, (−)-Phenylahistin, Myoseverin B,Resverastatin phosphate sodium.

In one aspect, compounds of Formula (I), Formula (II) and Formula (III)are co-administered with thrombolytic agents (e.g., alteplaseanistreplase, streptokinase, urokinase, or tissue plasminogenactivator), heparin, tinzaparin, warfarin, dabigatran (e.g., dabigatranetexilate), factor Xa inhibitors (e.g., fondaparinux, draparinux,rivaroxaban, DX-9065a, otamixaban, LY517717, or YM150), ticlopidine,clopidogrel, CS-747 (prasugrel, LY640315), ximelagatran, or BIBR 1048.

In some embodiments, compounds of Formula (I), Formula (II) and Formula(III) are used in combination with anti-emetic agents to treat nausea oremesis, which may result from the use of a compound of Formula (I),Formula (II) or Formula (III), anti-cancer agent(s) and/or radiationtherapy.

Anti-emetic agents include, but are not limited to: neurokinin-1receptor antagonists, 5HT3 receptor antagonists (such as ondansetron,granisetron, tropisetron, Palonosetron, and zatisetron), GABA_(B)receptor agonists (such as baclofen), corticosteroids (such asdexamethasone, prednisone, prednisolone, or others), dopamineantagonists (such as, but not limited to, domperidone, droperidol,haloperidol, chlorpromazine, promethazine, prochlorperazine,metoclopramide), antihistamines (H1 histamine receptor antagonists, suchas but not limited to, cyclizine, diphenhydramine, dimenhydrinate,meclizine, promethazine, hydroxyzine), cannabinoids (such as but notlimited to, cannabis, marinol, dronabinol), and others (such as, but notlimited to, trimethobenzamide; ginger, emetrol, propofol).

In some embodiments, compounds of Formula (I), Formula (II) and Formula(III) are used in combination with an agent useful in the treatment ofanemia. Such an anemia treatment agent is, for example, a continuouseythropoiesis receptor activator (such as epoetin-α).

In some embodiments, compounds of Formula (I), Formula (II) and Formula(III) are used in combination with an agent useful in the treatment ofneutropenia. Examples of agents useful in the treatment of neutropeniainclude, but are not limited to, a hematopoietic growth factor whichregulates the production and function of neutrophils such as a humangranulocyte colony stimulating factor, (G-CSF). Examples of a G-CSFinclude filgrastim.

In some embodiments, compounds of Formula (I), Formula (II) and Formula(III) are used in combination with radiation therapy (or radiotherapy).Radiation therapy is the treatment of cancer and other diseases withionizing radiation. Radiation therapy can be used to treat localizedsolid tumors, such as cancers of the skin, tongue, larynx, brain,breast, prostate, colon, uterus and/or cervix. It can also be used totreat leukemia and lymphoma (cancers of the blood-forming cells andlymphatic system, respectively).

A technique for delivering radiation to cancer cells is to placeradioactive implants directly in a tumor or body cavity. This is calledinternal radiotherapy (brachytherapy, interstitial irradiation, andintracavitary irradiation are types of internal radiotherapy.) Usinginternal radiotherapy, the radiation dose is concentrated in a smallarea, and the patient stays in the hospital for a few days. Internalradiotherapy is frequently used for cancers of the tongue, uterus,prostate, colon, and cervix.

The term “radiotherapy” or “ionizing radiation” include all forms ofradiation, including but not limited to α, β, and γ radiation andultraviolet light.

Immunosuppresants

In one aspect, compounds of Formula (I), Formula (II) and Formula (III)are to treat or reduce fibrosis in a mammal. In one aspect, compounds ofFormula (I), Formula (II) and Formula (III) are administered incombination with one or more immunosuppresants. Immunosuppressivetherapy is clinically used to treat or prevent the rejection oftransplanted organs and tissues (e.g. bone marrow, heart, kidney,liver); treatment of autoimmune diseases or diseases that are mostlikely of autoimmune origin (e.g. rheumatoid arthritis, myastheniagravis, systemic lupus erythematosus, Crohn's disease, and ulcerativecolitis); and treatment of some other non-autoimmune inflammatorydiseases (e.g. long term allergic asthma control), and in the treatmentof fibrotic conditions.

In some embodiments, a compound of Formula (I), Formula (II) or Formula(III) is administered with corticosteroids. In some embodiments, acompound of Formula (I), Formula (II) or Formula (III) is adminsiteredwith an a therapeutic agent selected from among: Calcineurin inhibitors(such as, but not limited to, cyclosporin, tacrolimus); mTOR inhibitors(such as, but not limited to, sirolimus, everolimus);anti-proliferatives (such as, but not limited to, azathioprine,mycophenolic acid); corticosteroids (such as, but not limited to,prednisone, cortisone acetate, prednisolone, methylprednisolone,dexamethasone, betamethasone, triamcinolone, beclometasone,fludrocortisone acetate, deoxycorticosterone acetate, aldosterone,hydrocortisone); antibodies (such as, but not limited to, monoclonalanti-IL-2Ra receptor antibodies (basiliximab, daclizumab), polyclonalanti-T-cell antibodies (anti-thymocyte globulin (ATG), anti-lymphocyteglobulin (ALG)), B-cell antagonists, rituximab, natalizumab.

Other therapeutic agents include, but are not limited to:cyclophosphamide, penicillamine, cyclosporine, nitrosoureas, cisplatin,carboplatin, oxaliplatin, methotrexate, azathioprine, mercaptopurine,pyrimidine analogues, protein synthesis inhibitors, dactinomycin,anthracyclines, mitomycin C, bleomycin, mithramycin, Atgam®,Thymoglobuline®, OKT3®, basiliximab, daclizumab, cyclosporin,tacrolimus, sirolimus, Interferons (IFN-β, IFN-γ), opioids, TNF bindingproteins (infliximab, etanercept, adalimumab, golimumab), leflunomide,gold thioglucose, gold thiomalate, aurofin, sulfasalazine,hydroxychloroquinine, minocycline, rapamicin, mycophenolic acid,mycophenolate mofetil, FTY720, as well as those listed in U.S. Pat. No.7,060,697.

In one embodiment, compounds of Formula (I), Formula (II) and Formula(III) is administered in combination with Cyclosporin A (CsA) ortacrolimus (FK506). In one embodiment, a compound of Formula (I),Formula (II) or Formula (III) is administered to a mammal in combinationwith an anti-inflammatory agent including, but not limited to,non-steroidal anti-inflammatory drugs (NSAIDs) and corticosteroids(glucocorticoids).

NSAIDs include, but are not limited to: aspirin, salicylic acid,gentisic acid, choline magnesium salicylate, choline salicylate, cholinemagnesium salicylate, choline salicylate, magnesium salicylate, sodiumsalicylate, diflunisal, carprofen, fenoprofen, fenoprofen calcium,flurobiprofen, ibuprofen, ketoprofen, nabutone, ketolorac, ketorolactromethamine, naproxen, oxaprozin, diclofenac, etodolac, indomethacin,sulindac, tolmetin, meclofenamate, meclofenamate sodium, mefenamic acid,piroxicam, meloxicam, COX-2 specific inhibitors (such as, but notlimited to, celecoxib, rofecoxib, valdecoxib, parecoxib, etoricoxib,lumiracoxib, CS-502, JTE-522, L-745,337 and NS398).

Corticosteroids, include, but are not limited to: betamethasone,prednisone, alclometasone, aldosterone, amcinonide, beclometasone,betamethasone, budesonide, ciclesonide, clobetasol, clobetasone,clocortolone, cloprednol, cortisone, cortivazol, deflazacort,deoxycorticosterone, desonide, desoximetasone, desoxycortone,dexamethasone, diflorasone, diflucortolone, difluprednate, fluclorolone,fludrocortisone, fludroxycortide, flumetasone, flunisolide, fluocinoloneacetonide, fluocinonide, fluocortin, fluocortolone, fluorometholone,fluperolone, fluprednidene, fluticasone, formocortal, halcinonide,halometasone, hydrocortisone/cortisol, hydrocortisone aceponate,hydrocortisone buteprate, hydrocortisone butyrate, loteprednol,medrysone, meprednisone, methylprednisolone, methylprednisoloneaceponate, mometasone furoate, paramethasone, prednicarbate,prednisone/prednisolone, rimexolone, tixocortol, triamcinolone, andulobetasol.

In one embodiment, compounds of Formula (I), Formula (II) and Formula(III) are administered in combination with leukotriene receptorantagonists including, but are not limited to, BAY u9773 (see EP00791576; published 27 Aug. 1997), DUO-LT (Tsuji et al, Org. Biomol.Chem., 1, 3139-3141, 2003), zafirlukast, montelukast, prankulast, andderivatives or analogs thereof.

Other Combination Therapies

In another embodiment described herein, methods for treatment ofLPA-dependent or LPA-mediated conditions or diseases, such asatherosclerosis, comprises administration to a patient compounds,pharmaceutical compositions, or medicaments described herein incombination with at least one additional agent selected, by way ofexample only, HMG-CoA reductase inhibitors (e.g., statins in theirlactonized or dihydroxy open acid forms and pharmaceutically acceptablesalts and esters thereof, including but not limited to lovastatin;simvastatin; dihydroxy open-acid simvastatin, particularly the ammoniumor calcium salts thereof; pravastatin, particularly the sodium saltthereof; fluvastatin, particularly the sodium salt thereof;atorvastatin, particularly the calcium salt thereof; nisvastatin, alsoreferred to as NK-104; rosuvastatin); agents that have bothlipid-altering effects and other pharmaceutical activities; HMG-CoAsynthase inhibitors; cholesterol absorption inhibitors such asezetimibe; cholesterol ester transfer protein (CETP) inhibitors, forexample JTT-705 and CP529, 414; squalene epoxidase inhibitors; squalenesynthetase inhibitors (also known as squalene synthase inhibitors);acyl-coenzyme A: cholesterol acyltransferase (ACAT) inhibitors includingselective inhibitors of ACAT-1 or ACAT-2 as well as dual inhibitors ofACAT-1 and -2; microsomal triglyceride transfer protein (MTP)inhibitors; probucol; niacin; bile acid sequestrants; LDL (low densitylipoprotein) receptor inducers; platelet aggregation inhibitors, forexample glycoprotein IIb/IIIa fibrinogen receptor antagonists andaspirin; human peroxisome proliferator activated receptor gamma (PPARγ)agonists, including the compounds commonly referred to as glitazones,for example troglitazone, pioglitazone and rosiglitazone and includingthose compounds included within the structural class known asthiazolidinediones as well as those PPARγ agonists outside thethiazolidinedione structural class; PPARα agonists such as clofibrate,fenofibrate including micronized fenofibrate, and gemfibrozil; PPAR dualα/γ agonists such as5-[(2,4-dioxo-5-thiazolidinyl)methyl]-2-methoxy-N-[[4-(trifluoromethyl)phenyl]methyl]-benzamide,known as KRP-297; vitamin B6 (also known as pyridoxine) and thepharmaceutically acceptable salts thereof such as the HCl salt; vitaminB12 (also known as cyanocobalamin); folic acid or a pharmaceuticallyacceptable salt or ester thereof such as the sodium salt and themethylglucamine salt; anti-oxidant vitamins such as vitamin C and E andbeta carotene; beta-blockers; angiotensin II antagonists such aslosartan; angiotensin converting enzyme inhibitors such as enalapril andcaptopril; calcium channel blockers such as nifedipine and diltiazam;endothelian antagonists; agents that enhance ABC1 gene expression; FXRand LXR ligands including both inhibitors and agonists; bisphosphonatecompounds such as alendronate sodium; and cyclooxygenase-2 inhibitorssuch as rofecoxib and celecoxib.

In another embodiment described herein, methods for treatment ofLPA-dependent or LPA-mediated conditions or diseases, such as thetherapy of stroke, comprises administration to a patient compounds,pharmaceutical compositions, or medicaments described herein incombination with at least one additional agent selected from, by way ofexample only, COX-2 inhibitors; nitric oxide synthase inhibitors, suchas N-(3-(aminomethyl)benzyl)acetamidine; Rho kinase inhibitors, such asfasudil; angiotension II type-1 receptor antagonists, includingcandesartan, losartan, irbesartan, eprosartan, telmisartan andvalsartan; glycogen synthase kinase 3 inhibitors; sodium or calciumchannel blockers, including crobenetine; p38 MAP kinase inhibitors,including SKB 239063; thromboxane AX-synthetase inhibitors, includingisbogrel, ozagrel, ridogrel and dazoxiben; statins (HMG CoA reductaseinhibitors), including lovastatin, simvastatin, dihydroxy open-acidsimvastatin, pravastatin, fluvastatin, atorvastatin, nisvastatin, androsuvastatin; neuroprotectants, including free radical scavengers,calcium channel blockers, excitatory amino acid antagonists, growthfactors, antioxidants, such as edaravone, vitamin C, TROLOX™, citicolineand minicycline, and reactive astrocyte inhibitors, such as(2R)-2-propyloctanoic acid; beta andrenergic blockers, such aspropranolol, nadolol, timolol, pindolol, labetalol, metoprolol,atenolol, esmolol and acebutolol; NMDA receptor antagonists, includingmemantine; NR2B antagonists, such as traxoprodil; 5-HT1A agonists;receptor platelet fibrinogen receptor antagonists, including tirofibanand lamifiban; thrombin inhibitors; antithrombotics, such as argatroban;antihypertensive agents, such as enalapril; vasodilators, such ascyclandelate; nociceptin antagonists; DPIV antagonists; GABA 5 inverseagonists; and selective androgen receptor modulators.

In another embodiment described herein, methods for treatment ofLPA-dependent or LPA-mediated conditions or diseases, such as thetherapy of interstitial cystitis, comprises administration to a patientcompounds, pharmaceutical compositions, or medicaments described hereinin combination with at least one additional agent selected from, by wayof example only, dimethylsulfoxide, omalizumab, and pentosanpolysulfate.

In yet another embodiment described herein, methods for treatingLPA-dependent or LPA-mediated conditions or diseases, such as thetherapy of respiratory disorders (e.g., asthma, COPD and rhinitis),comprises administration to a patient compounds, pharmaceuticalcompositions, or medicaments described herein in combination with atleast one agent used in the treatment of respiratory conditions. Agentsused in the treatment of respiratory conditions include, but are notlimited to, bronchodilators (e.g., sympathomimetic agents and xanthinederivatives), leukotriene receptor antagonists, leukotriene formationinhibitors, leukotriene modulators, nasal decongestants, respiratoryenzymes, lung surfactants, antihistamines (e.g., mepyramine(pyrilamine), antazoline, diphenhydramine, carbinoxamine, doxylamine,clemastine, dimenhydrinate, pheniramine, chlorphenamine(chlorpheniramine), dexchlorpheniramine, brompheniramine, triprolidine,cetirizine, cyclizine, chlorcyclizine, hydroxyzine, meclizine,loratadine, desloratidine, promethazine, alimemazine (trimeprazine),cyproheptadine, azatadine, ketotifen, acrivastine, astemizole,cetirizine, mizolastine, terfenadine, azelastine, levocabastine,olopatadine, levocetirizine, fexofenadine), mucolytics, corticosteroids,anticholinergics, antitussives, analgesics, expectorants, albuterol,ephedrine, epinephrine, fomoterol, metaproterenol, terbutaline,budesonide, ciclesonide, dexamethasone, flunisolide, fluticasonepropionate, triamcinolone acetonide, ipratropium bromide,pseudoephedrine, theophylline, montelukast, zafirlukast, ambrisentan,bosentan, enrasentan, sitaxsentan, tezosentan, iloprost, treprostinil,pirfenidone, 5-lipoxygenase-activating protein (FLAP) inhibitors, FLAPmodulators and 5-LO inhibitors.

In a specific embodiment described herein, methods for treatingLPA-dependent or LPA-mediated conditions or diseases, such as thetherapy of asthma and/or COPD, comprises administration to a patientanti-inflammatory agents. In certain embodiments, methods for treatingLPA-dependent or LPA-mediated conditions or diseases, such as thetherapy of asthma and/or COPD, comprise administration to a patientcompounds, pharmaceutical compositions, or medicaments described hereinin combination with at least one additional agent selected from, but notlimited to, epinephrine, isoproterenol, orciprenaline, bronchodilators,glucocorticoids, leukotriene modifiers, mast-cell stabilizers,xanthines, anticholinergics, β-2 agonists, FLAP inhibitors, FLAPmodulators or 5-LO inhibitors. β-2 agonists include, but are not limitedto, short-acting β-2 agonists (e.g., salbutamol (albuterol),levalbuterol, terbutaline, pirbuterol, procaterol, metaproterenol,fenoterol and bitolterol mesylate) and long-acting β-2 agonists (e.g.,salmeterol, formoterol, bambuterol and clenbuterol). FLAP inhibitorsand/or FLAP modulators include, but are not limited to,3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionicacid,3-[3-tert-butylsulfanyl-1-[4-(6-ethoxy-pyridin-3-yl)-benzyl]-5-(5-methyl-pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionicacid, MK-886, MK-0591, BAY-x1005 and compounds found in US 2007/0225285,US 2007/0219206, US 2007/0173508, US 2007/0123522 and US 2007/0105866(each of which are hereby incorporated by reference). Glucocorticoidsinclude, but are not limited to, beclometasone, budesonide, ciclesonide,fluticasone and mometasone. Anticholinergics include, but are notlimited to, ipratropium and tiotropium. Mast cell stabilizers include,but are not limited to, cromoglicate and nedocromil. Xanthines include,but are not limited to, amminophylline, theobromine and theophylline.Leukotriene antagonists include, but are not limited to, montelukast,tomelukast, pranlukast and zafirlukast. 5-LO inhibitors include, but arenot limited to, zileuton, VIA-2291 (ABT761), AZ-4407 and ZD-2138 andcompounds found in US 2007/0149579, WO2007/016784.

In another specific embodiment described herein, methods for treatingLPA-dependent or LPA-mediated conditions or diseases, such as thetherapy of allergic diseases or conditions, comprises administration toa patient compounds, pharmaceutical compositions, or medicamentsdescribed herein in combination with at least one additional agentselected from, by way of example only, antihistamines, leukotrieneantagonists, corticosteroids and decongestants. Leukotriene antagonistsinclude, but are not limited to, montelukast, tomelukast, pranlukast andzafirlukast.

In one aspect, LPA receptor antagonists described herein are admisteredin combination with one or more agents used to treat used to treatasthma, including, but not limited to: combination inhalers (fluticasoneand salmeterol oral inhalation (e.g. Advair)); inhaled Beta-2 agonists(albuterol inhaler; albuterol nebulizer solution; formoterol;isoproterenol oral inhalation; levalbuterol; metaproterenol inhalation;pirbuterol acetate oral inhalation; salmeterol aerosol inhalation;salmeterol powder inhalation; terbutaline inhaler); inhaledcorticosteroids (beclomethasone oral inhalation; budesonide inhalationsolution; budesonide inhaler; flunisolide oral inhalation; fluticasoneinhalation aerosol; fluticasone powder for oral inhalation; mometasoneinhalation powder; triamcinolone oral inhalation); leukotriene modifiers(montelukast; zafirlukast; zileuton); mast cell stabilizers (cromolyninhaler; nedocromil oral inhalation); monoclonal antibodies(omalizumab); oral Beta-2 agonists (albuterol oral syrup; albuterol oraltablets; metaproterenol; terbutaline); bronchodilator (aminophylline;oxtriphylline; theophylline).

In one aspect, LPA receptor anatogonists described herein are admisteredin combination with one or more agents used to treat allergy, including,but not limited to: antihistamine and decongestant combinations(cetirizine and pseudoephedrine; desloratadine and pseudoephedrine ER;fexofenadine and pseudoephedrine; loratadine and pseudoephedrine);antihistamines (azelastine nasal spray; brompheniramine; brompheniramineoral suspension; carbinoxamine; cetirizine; chlorpheniramine;clemastine; desloratadine; dexchlorpheniramine ER; dexchlorpheniramineoral syrup; diphenhydramine oral; fexofenadine; loratadine;promethazine); decongestants (pseudoephedrine); leukotriene modifiers(montelukast; montelukast granules); nasal anticholinergics(ipratropium); nasal corticosteroids (beclomethasone nasal inhalation;budesonide nasal inhaler; flunisolide nasal inhalation; fluticasonenasal inhalation; mometasone nasal spray; triamcinolone nasalinhalation; triamcinolone nasal spray); nasal decongestants(phenylephrine); nasal mast cell stabilizers (cromolyn nasal spray).

In one aspect, LPA receptor anatogonists described herein are admisteredin combination with one or more agents used to treat chronic obstructivepulmonary disease (COPD), including, but not limited to:anticholinergics—ipratropium bromide oral inhalation); combinationInhalers (albuterol and ipratropium (e.g. Combivent, DuoNeb);fluticasone and salmeterol oral inhalation (e.g. Advair));corticosteroids (dexamethasone tablets; fludrocortisone acetate;hydrocortisone tablets; methylprednisolone; prednisolone liquid;prednisone oral; triamcinolone oral); inhaled Beta-2 Agonists (albuterolinhaler; albuterol nebulizer solution; formoterol; isoproterenol oralinhalation; levalbuterol; metaproterenol inhalation; pirbuterol acetateoral inhalation; salmeterol aerosol inhalation; salmeterol powderinhalation; terbutaline inhaler); inhaled Corticosteroids(beclomethasone oral inhalation; budesonide inhalation solution;budesonide inhaler; flunisolide oral inhalation; fluticasone inhalationaerosol; fluticasone powder for oral inhalation; triamcinolone oralinhalation); mukolytics (guaifenesin); oral Beta-2 agonists (albuteroloral syrup; albuterol oral tablets; metaproterenol; terbutaline);bronchodilator (aminophylline; oxtriphylline; theophylline).

In one embodiment, a compound of Formula (I), Formula (II) or Formula(III) described herein is administered to a patient in combination withinhaled corticosteroids.

In one embodiment, a compound of Formula (I), Formula (II) or Formula(III) described herein is administered to a patient in combination withbeta2-adrenergic receptor agonists. In one embodiment, a compound ofFormula (I), Formula (II) or Formula (III) is administered to a patientin combination with short acting beta2-adrenergic receptor agonists. Inone embodiment, a compound of Formula (I), Formula (II) or Formula (III)is administered to a patient in combination with long-actingbeta2-adrenergic receptor agonists.

Kits/Articles of Manufacture

For use in the therapeutic applications described herein, kits andarticles of manufacture are also described herein. Such kits cancomprise a carrier, package, or container that is compartmentalized toreceive one or more containers such as vials, tubes, and the like, eachof the container(s) comprising one of the separate elements to be usedin a method described herein. Suitable containers include, for example,bottles, vials, syringes, and test tubes. The containers are formed fromany acceptable material including, e.g., glass or plastic.

For example, the container(s) can comprise one or more compoundsdescribed herein, optionally in a composition or in combination withanother agent as disclosed herein. The container(s) optionally have asterile access port (for example the container can be an intravenoussolution bag or a vial having a stopper pierceable by a hypodermicinjection needle). Such kits optionally comprising a compound with anidentifying description or label or instructions relating to its use inthe methods described herein.

A kit will typically comprise one or more additional containers, eachwith one or more of various materials (such as reagents, optionally inconcentrated form, and/or devices) desirable from a commercial and userstandpoint for use of a compound described herein. Non-limiting examplesof such materials include, but not limited to, buffers, diluents,filters, needles, syringes; carrier, package, container, vial and/ortube labels listing contents and/or instructions for use, and packageinserts with instructions for use. A set of instructions will alsotypically be included.

A label can be on or associated with the container. A label can be on acontainer when letters, numbers or other characters forming the labelare attached, molded or etched into the container itself; a label can beassociated with a container when it is present within a receptacle orcarrier that also holds the container, e.g., as a package insert. Alabel can be used to indicate that the contents are to be used for aspecific therapeutic application. The label can also indicate directionsfor use of the contents, such as in the methods described herein.

EXAMPLES

These examples are provided for illustrative purposes only and not tolimit the scope of the claims provided herein.

Synthesis of Compounds

(R)-2′-chloro-alpha-methylbenzyl alcohol

Using the procedure of Meier et at (Tetrahedron, 1996, 52, 589; Method3), 2′-chloroacetophenone (Aldrich) is reduced to give(R)-2′-chloro-alpha-methylbenzylalcohol. (% e.e. is determined by HPLCanalysis of the acetate derivative (by reacting the benzyl alcohol withacetyl chloride and triethylamne in methylene chloride) using ChiralcelOD eluted with 99:1 Hexane:Ethanol)

(S)-2′-chloro-alpha-methylbenzyl alcohol

Using the procedure of Meier et at (Tetrahedron, 1996, 52, 589; Method3), 2′-chloroacetophenone (Aldrich) is reduced to give(S)-2′-chloro-alpha-methylbenzylalcohol. (% e.e. is determined by HPLCanalysis of the acetate derivative (by reacting the benzyl alcohol withacetyl chloride and triethylamne in methylene chloride) using ChiralcelOD eluted with 99:1 Hexane:Ethanol)

(R)-2′-fluoro-alpha-methylbenzyl alcohol

Using the procedure of Meier et at (Tetrahedron, 1996, 52, 589; Method3), 2′-fluoroacetophenone (Aldrich) is reduced to give(R)-2′-fluoro-alpha-methylbenzylalcohol. (% e.e. is determined by HPLCanalysis of the acetate derivative (by reacting the benzyl alcohol withacetyl chloride and triethylamne in methylene chloride) using ChiralcelOD eluted with 99.8:0.2 Hexane:Ethanol).

(S)-2′-fluoro-alpha-methylbenzyl alcohol

Using the procedure of Meier et at (Tetrahedron, 1996, 52, 589; Method3), 2′-fluoroacetophenone (Aldrich) was reduced to give(S)-2′-fluoro-alpha-methylbenzylalcohol. (% e.e. is determined by HPLCanalysis of the acetate derivative (by reacting the benzyl alcohol withacetyl chloride and triethylamne in methylene chloride) using ChiralcelOD eluted with 99.8:0.2 Hexane:Ethanol).

Example 1 Synthesis of5-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-pent-4-ynoicacid (Compound 1-1)

Step 1: 3-Methylamino-but-2-enoic acid methyl ester

To a solution of methyl acetoacetate (29.4 g, 253 mmol) in MeOH (30 mL)was added methylamine (33 wt % in EtOH; 48 mL, 385 mmol) dropwise atroom temperature. The reaction was stirred for 1 hour, and thenconcentrated and dried to give the title compound as a white crystallinesolid.

Step 2: 2-(4-Bromo-benzoyl)-3-[(E)-methylimino]-butyric acid methylester

4-Bromobenzoyl chloride (17.14 g, 78.1 mmol) and3-methylamino-but-2-enoic acid methyl ester (10.0 g, 78.1 mmol) werecombined in THF (600 mL) and cooled to 0° C. Pyridine (6.8 mL, 85.9mmol) was added by syringe, and the reaction was stirred overnight atroom temperature. Solid pyridine hydrochloride was observed on the flaskwalls, so the solution was decanted and then concentrated to 10% of itsoriginal volume. The residue was diluted with EtOAc (400 mL) and washedtwice with H₂O. The organic layer was dried, filtered, and concentratedto give the title compound.

Step 3: 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid methylester

2-(4-Bromo-benzoyl)-3-[(E)-methylimino]-butyric acid methyl ester (23.81g, 76.6 mmol) and hydroxylamine hydrochloride (5.32 g, 76.6 mmol) werecombined in acetic acid (400 mL) and stirred at 60° C. overnight. Aftercooling to room temperature, the mixture was concentrated to dryness,and then concentrated twice from toluene (200 mL) to remove residualacetic acid. The residue was partitioned between EtOAc and H₂O, and theorganic layer was separated and washed twice with H₂O. The combinedaqueous layers were back-extracted with EtOAc, and the combined organiclayers were concentrated. The crude material was filtered through a plugof silica gel, and the filtrate was concentrated to give the titlecompound.

Step 4: 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid

5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid methyl ester(7.5 g, 25 mmol) was suspended in 2:1 methanol:water. Lithium hydroxide(2.1 g, 50 mmol) was added, and the reaction was stirred overnight atroom temperature. The mixture was acidified to pH 1 with 1N aqueous HCland extracted with EtOAc. The combined organic layers were dried overMgSO₄, filtered, and concentrated to give the title compound.

Step 5: [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-chloro-phenyl)-ethyl ester

To a suspension of 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylicacid (7.0 g, 24.8 mmol) and triethylamine (3.46 g, 24.8 mmol) in toluene(250 mL) was added diphenylphosphoryl azide (5.31 mL, 24.8 mmol),followed by 2-chloro-alpha-methylbenzyl alcohol (3.88 g, 24.8 mmol), andthe reaction was stirred at 80° C. for 2.5 hours. The mixture waspartitioned between EtOAc and H₂O, and the aqueous layer was extractedwith EtOAc. The combined organic layers were dried over MgSO₄, filtered,and concentrated, and the residue was dissolved in EtOAc and filteredthrough Celite to remove a precipitate that was present. The crudematerial was purified by silica gel chromatography to give the titlecompound.

Step 6:5-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-pent-4-ynoicacid

[5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-chloro-phenyl)-ethyl ester (0.200 g, 0.46 mmol), 4-pentynoic acid(0.054 g, 0.55 mmol), copper iodide (spatula tip), triethylamine (0.12mL, 0.84 mmol), and tetrakis(triphenylphosphine)palladium(0) (0.053 g,0.05 mmol) were combined in DME (5 mL), and the reaction was purged withN₂ for 15 minutes and then stirred at 80° C. overnight. The mixture waspartitioned between EtOAc and H₂O, and the aqueous layer was separatedand extracted with EtOAc. The combined organic layers were dried overMgSO₄, filtered, and concentrated, and the residue was purified bypreparative HPLC to give the title compound. Mass spec. data (M+H)=453.

Example 2 Synthesis of6-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-hex-5-ynoicacid (Compound 1-2)

Step 1:6-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-hex-5-ynoicacid

To a solution of [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamicacid 1-(2-chloro-phenyl)-ethyl ester (0.200 g, 0.46 mmol) and 5-hexynoicacid (0.06 mL, 0.55 mmol) in DME (5 mL) was added copper iodide (0.017g, 0.09 mmol), followed by tetrakis(triphenylphosphine)palladium(0)(0.032 g, 0.05 mmol), and the mixture was purged with N₂ for 5 minutes.Triethylamine (0.26 mL, 1.83 mmol) was added, and the reaction waspurged with N₂ for another 10 minutes and then stirred at 80° C. for 2hours. The mixture was partitioned between EtOAc and H₂O, and theaqueous layer was separated and extracted with EtOAc. The combinedorganic layers were dried over MgSO₄, filtered, and concentrated, andthe residue was purified by preparative HPLC to give the title compound.

Mass spec. data (M+H)=467.

Example 3 Synthesis of7-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-hept-6-ynoicacid (Compound 1-3)

Step 1:7-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-hept-6-ynoicacid

Prepared according to the procedure described in Example 2, Step 1,using the following starting materials:[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-chloro-phenyl)-ethyl ester and 6-heptynoic acid. Mass spec. data(M+H)=481.

Example 4 Synthesis of5-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-2,2-dimethyl-pent-4-ynoicacid (Compound 1-4)

Step 1: 2,2-Dimethyl-pent-4-ynoic acid ethyl ester

Diisopropylamine (45.9 mL, 0.33 mol) was dissolved in THF (300 mL) andcooled to 0° C. under N₂. n-Butyllithium (2.5M in hexanes; 131.0 mL,0.33 mol) was added dropwise over 30 minutes, and the mixture wasstirred at 0° C. for an additional 30 minutes and then cooled to −78° C.Ethyl isobutyrate (40 mL, 0.30 mol) in THF (30 mL) was added dropwise,and the solution was stirred at −78° C. for 1 hour. Propargyl bromide(36.4 mL, 0.33 moml) in HMPA (60 mL) was added dropwise, and thereaction was stirred at −78° C. for 1 hour and then quenched withsaturated aqueous NH₄Cl and warmed to room temperature. THF was removedin vacuo, and the residue was dissolved in Et₂O and washed four timeswith H₂O. The organic layer was dried over MgSO₄, filtered, andconcentrated, and the crude material was distilled under reducedpressure (˜29 inHg, by 68-70° C.) to give the title compound.

Step 2:5-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-2,2-dimethyl-pent-4-ynoicacid ethyl ester

Prepared according to the procedure described in Example 2, Step 1,using the following starting materials:[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-chloro-phenyl)-ethyl ester and 2,2-dimethyl-pent-4-ynoic acid; nopurification was performed.

Step 2:5-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-2,2-dimethyl-pent-4-ynoicacid

To5-(4-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-2,2-dimethyl-pent-4-ynoicacid ethyl ester (0.46 mmol) in MeOH and H₂O was added lithiumhydroxide, and the reaction was stirred at 60° C. overnight. The mixturewas acidified and extracted with EtOAc. The combined organic layers weredried over MgSO₄, filtered, and concentrated, and the residue waspurified by preparative HPLC to give the title compound. Mass spec. data(M+H)=481.

Example 5 Synthesis of1-[3-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-5-trifluoromethyl-1H-pyrazole-4-carboxylicacid (Compound 2-1)

Step 1:{5-[4-(3-Hydroxy-prop-1-ynyl)-phenyl]-3-methyl-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester

Prepared according to the procedure described in Example 2, Step 1,using the following starting materials:[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-chloro-phenyl)-ethyl ester and propargy alcohol; purification bysilica gel chromatography was utilized.

Step 2:1-[3-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-5-trifluoromethyl-1H-pyrazole-4-carboxylicacid ethyl ester

To a solution of{5-[4-(3-hydroxy-prop-1-ynyl)-phenyl]-3-methyl-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester (0.200 g, 0.49 mmol), ethyl5-(trifluoromethyl)-1H-pyrazole-4-carboxylate (0.101 g, 0.49 mmol), andtriphenylphosphine (0.127 g, 0.49 mmol) in THF (4 mL) was addeddiisopropyl azodicarboxylate (0.10 mL, 0.49 mmol), and the reaction wasstirred at room temperature for 2 hours. The mixture was concentrated,and the residue was purified by preparative HPLC to give the titlecompound.

Step 3:1-[3-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-5-trifluoromethyl-1H-pyrazole-4-carboxylicacid

To a solution of1-[3-(4-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-5-trifluoromethyl-1H-pyrazole-4-carboxylicacid ethyl ester (0.155 g, 0.26 mmol) in 2:1 MeOH:H₂O (3 mL) was addedlithium hydroxide (0.033 g, 0.79 mmol), and the reaction was stirredovernight at room temperature. The mixture was acidified with 1N aqueousHCl and extracted with EtOAc, and the combined organic layers weredried, filtered, and concentrated. The residue was purified bypreparative HPLC to give the title compound. Mass spec. data (M+H)=573.

Example 6 Synthesis of1-[3-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-1H-pyrazole-4-carboxylicacid (Compound 2-2)

Step 1:1-[3-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-1H-pyrazole-4-carboxylicacid ethyl ester

Prepared according to the procedure described in Example 5, Step 2,using the following starting materials:{5-[4-(3-hydroxy-prop-1-ynyl)-phenyl]-3-methyl-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester and ethyl 4-pyrazolecarboxylate.

Step 2:1-[3-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-1H-pyrazole-4-carboxylicacid

Prepared according to the procedure described in Example 5, Step 3,using the following starting material:1-[3-(4-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-1H-pyrazole-4-carboxylicacid ethyl ester.

Mass spec. data (M+H)=505.

Example 7 Synthesis of1-[3-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-1H-imidazole-4-carboxylicacid (Compound 2-3)

Step 1:1-[3-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-1H-imidazole-4-carboxylicacid methyl ester

Prepared according to the procedure described in Example 5, Step 2,using the following starting materials:{5-[4-(3-hydroxy-prop-1-ynyl)-phenyl]-3-methyl-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester and methyl 4-imidazolecarboxylate.

Step 2:1-[3-(4-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-1H-imidazole-4-carboxylicacid

Prepared according to the procedure described in Example 5, Step 3,using the following starting material:1-[3-(4-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-phenyl)-prop-2-ynyl]-1H-imidazole-4-carboxylicacid methyl ester.

Mass spec. data (M+H)=505.

In vitro Assay(s)

Example 9 Establishment of a CHO Cell Line Stably Expressing Human LPA₁

A 1.1 kb cDNA encoding the human LPA₁ receptor was cloned from humanlung. Human lung RNA (Clontech Laboratories, Inc. USA) was reversetranscribed using the RETROscript kit (Ambion, Inc.) and the full-lengthcDNA for human LPA₁ was obtained by PCR of the reverse transcriptionreaction. The nucleotide sequence of the cloned human LPA₁ wasdetermined by sequencing and confirmed to be identical to the publishedhuman LPA₁ sequence (An et al. Biochem. Biophys. Res. Commun. 231:619(1997). The cDNA was cloned into the pCDNA5/FRT expression plasmid andtransfected in CHO cells using lipofectamine 2000 (Invitrogen Corp.,USA). Clones stably expressing human LPA₁ were selected using hygromycinand identified as cells that show Ca-influx in response to LPA.

Example 10 Generation of Cells Transiently Expressing Human LPA₂

A vector containing the human LPA₂ receptor cDNA was obtained from theMissouri S&T cDNA Resource Center (www.cdna.org). The full-length cDNAfragment for human LPA₂ was obtained by PCR from the vector. Thenucleotide sequence of the cloned human LPA₂ was determined bysequencing and confirmed to be identical to the published human LPA₂sequence (NCBI accession number NM_(—)004720). The cDNA was cloned intothe pCDNA3.1 expression plasmid and transfected into B103 cells(Invitrogen Corp., USA) by seeding cells in a 96-well poly-D-lysinecoated plate at 30,000-35,000 cells per well together with 0.2 μllipofectamine 2000 and 0.2 μg of the LPA₂ expression vector. Cells werecultured overnight in complete media before being assayed forLPA-induced Ca-influx.

Example 11 Establishment of a CHO Cell Line Stably Expressing Human LPA₃

A vector containing the human LPA₃ receptor cDNA was obtained from theMissouri S&T cDNA Resource Center (www.cdna.org). The full-length cDNAfragment for human LPA₃ was obtained by PCR from the vector. Thenucleotide sequence of the cloned human LPA₃ was determined bysequencing and confirmed to be identical to the published human LPA₃sequence (NCBI accession number NM_(—)012152). The cDNA was cloned intothe pCDNA5/FRT expression plasmid and transfected in CHO cells usinglipofectamine 2000 (Invitrogen Corp., USA). Clones stably expressinghuman LPA₃ were selected using hygromycin and identified as cells thatshow Ca-influx in response to LPA.

Example 12 LPA1 and LPA3 Calcium Flux Assays

Human LPA₁ or LPA₃ expressing CHO cells are seeded at 20,000-45,000cells per well in a 96-well poly-D-lysine coated plate one or two daysbefore the assay. Prior to the assay, the cells are washed once with PBSand then cultured in serum-free media overnight. On the day of theassay, a calcium indicator dye (Calcium 4, Molecular Devices) in assaybuffer (HBSS with Ca²⁺ and Mg²⁺ and containing 20 mM Hepes and 0.3%fatty-acid free human serum albumin) is added to each well andincubation continued for 1 hour at 37° C. 10 μl of test compounds in2.5% DMSO are added to the cells and incubation continued at roomtemperature for 30 minutes. Cells are the stimulated by the addition of10 nM LPA and intracellular Ca²⁺ measured using the Flexstation 3(Molecular Devices). IC₅₀s are determined using Graphpad prism analysisof drug titration curves.

Example 13 LPA2 Calcium Flux Assay

Following an overnight culture with lipofectamine 2000 and the LPA₂expression vector, the B103 cells are washed once with PBS then serumstarved for 4 hours. A calcium indicator dye (Calcium 4, MolecularDevices) in assay buffer (HBSS with Ca²⁺ and Mg²⁺ and containing 20 mMHepes and 0.3% fatty-acid free human serum albumin) is added to eachwell and incubation continued for 1 hour at 37° C. 10 μl of testcompounds in 2.5% DMSO are added to the cells and incubation continuedat room temperature for 30 minutes. Cells are the stimulated by theaddition of 10 nM LPA and intracellular Ca²⁺ measured using theFlexstation 3 (Molecular Devices). IC₅₀s are determined using Graphpadprism analysis of drug titration curves.

Example 14 GTPγS Binding Assay

The ability of a compound to inhibit binding of GTP to LPA₁ is assessedvia a membrane GTPγS assay. CHO cells stably expressing the recombinanthuman LPA₁ receptor are resuspended in 10 mM Hepes, 7.4 containing 1 mMDTT, lysed and centrifuged at 75,000×g to pellet the membranes. Themembranes are resuspended in 10 mM Hepes, 7.4 containing 1 mM DTT and10% glycerol. Membranes (˜25 μg per well) are incubated in 96-wellplates with 0.1 nM [³⁵S]-GTPγS, 900 nM LPA, 5 μM GDP, and test compoundin Assay Buffer (50 mM Hepes, pH 7.4, 100 mM NaCl, 10 mM MgCl₂, 50 μg/mlsaponin and 0.2% fatty-acid free human serum albumin) for 30 minutes at30° C. The reactions are terminated by rapid filtration through WhatmanGF/B glass fibre filter plates. The filter plates are washed 3 timeswith 1 ml cold Wash Buffer (50 mM Hepes, 7.5, 100 mM NaCl and 10 mMMgCl₂) and dried. Scintillant is then added to the plates and theradioactivity retained on the filters is determined on a PackardTopCount (Perkin Elmer). Specific binding is determined as totalradioactive binding minus non-specific binding in the absence of theligand (900 nM LPA). IC₅₀s were determined using Graphpad prism analysisof drug titration curves.

Illustrative in vitro biological data for representative compounds ofFormula (I) is presented in the Table below. Unless otherwise noted,compounds that were tested had an IC₅₀ of less than 50 μM in the HLPA₁Ca Flux assay.

HLPA₁ Ca Flux Compound Number IC₅₀ (μM) Compound 1-1 A Compound 1-2 ACompound 1-3 A Compound 1-4 A Compound 2-1 C Compound 2-2 B Compound 2-3A Compound 3-1 A A = less than 0.3 μM; B = greater than 0.3 μM and lessthan 1 μM; C = greater than 1 μM.

Example 15 LPA1 Chemotaxis Assay

Chemotaxis of the A2058 human melanoma cells was measured using theNeuroprobe ChemoTx® System plates (8 μm pore size, 5.7 mm diametersites). The filter sites were coated with 0.001% fibronectin (Sigma) in20 mM Hepes, pH 7.4 and allowed to dry. A2058 cells were serum-starvedfor 24 hours, then harvested with Cell Stripper and resuspended in DMEMcontaining 0.1% fatty-acid-free bovine serum albumin (BSA) to aconcentration of 1×10⁶/ml. Cells were mixed with an equal volume of testcompound (2×) in DMEM containing 0.1% fatty-acid-free BSA and incubatedat 37° C. for 15 minutes. LPA (100 nM in DMEM containing 0.1%fatty-acid-free BSA) or vehicle was added to each well of the lowerchamber and 50 μl of the cell suspension/test compound mix was appliedto the upper portion of the ChemoTx plate. Plates were incubated at 37°C. for three hours and then the cells removed from the upper portion byrinsing with PBS and scraping. The filter was dried then stained withHEMA 3 Staining System (Fisher Scientific). The absorbance of the filterwas read at 590 nM and IC₅₀s were determined using Symyx Assay Explorer.

In vivo Assay(s)

Example 16 Bleomycin-Induced Lung Fibrosis Model in Mice

Female CD-1 mice (Harlan, 25-30 g) are housed 4 per cage, given freeaccess to food and water and allowed to acclimate for at least 7 daysprior to test initiation. After the habituation phase, mice are lightlyanesthetized with isoflurane (5% in 100% O₂) and administered withbleomycin sulfate (0.01-5 U/kg, Henry Schein) via intratrachealinstillation (Cuzzocrea S et al. Am J Physiol Lung Cell Mol Physiol.2007 May; 292(5):L1095-104. Epub 2007 Jan. 12). Mice are returned totheir cages and monitored daily for the duration of the experiment. Testcompound or vehicle is delivered po, ip or sc daily. The route andfrequency of dosing is based on previously determined pharmacokineticproperties. All animals are sacrificed using inhaled isoflurane 3, 7,14, 21 or 28 days after bleomycin instillation. Following sacrifice,mice are intubated with a 20 gauge angiocatheter attached to a 1 mlsyringe. Lungs are lavaged with saline to obtain bronchoalveolar lavagefluid (BALF) and then removed and fixed in 10% neutral buffered formalinfor subsequent histopathological analysis. BALF is centrifuged for 10min at 800×g to pellet the cells and the cell supernatant removed andfrozen at −80° C. for subsequent protein analysis using the DC proteinassay kit (Biorad, Hercules, Calif.) and soluble collagen analysis usingSircol (Biocolor Ltd, UK). BALF is analyzed for concentrations ofinflammatory, pro-fibrotic and tissue injury biomarkers includingtransforming growth factor β1, hyaluronic acid, tissue inhibitor ofmetalloproteinase-1, matrix matelloproteinase-7, connective tissuegrowth factor and lactate dehydrogenase activity, using commerciallyavailable ELISA. The cell pellet is re-suspended in PBS. Total cellcounts are then obtained using a Hemavet hematology system (DrewScientific, Wayne, Pa.) and differential cells counts are determinedusing Shandon cytospin (Thermo Scientific, Waltham, Mass.). Lung tissueis stained using hematoxylin and eosin (H&E) and trichrome and lungfibrosis is determined by semiquantitative histopathological scoring(Ashcroft T. et al. J. Clin. Path. 1988; 41; 4, 467-470) using lightmicroscopy (10× magnification) and quantitative, computer-assisteddensitometry of collagen in lung tissue sections using light microscopy.The data are plotted using Graphpad prism and statistical differencesbetween groups determined.

Example 17 Mouse Carbon Tetrachloride (CCl₄)-Induced Liver FibrosisModel

Female C57BL/6 mice (Harlan, 20-25 g) housed 4/cage are given freeaccess to food and water and allowed to acclimate for at least 7 daysprior to test initiation. After the habituation phase, mice receive CCl₄(0.5-1.0 ml/kg body weight) diluted in corn oil vehicle (100 μL volume)via i.p. injection twice a week for 4-6 weeks. (Higazi, A. A. et al.,Clin Exp Immunol. 2008 April; 152(1):163-73. Epub 2008 Feb. 14). Controlmice receive an equivalent volume of corn oil vehicle only. Testcompound or vehicle is delivered po, ip or sc daily. At the end of thestudy (4-6 weeks after first i.p. injection of CCl₄), mice aresacrificed using inhaled isoflurane and blood is drawn via cardiacpuncture for subsequent analysis of ALT/AST levels. The liver isharvested, and one half of the liver is frozen at −80° C. and the otherhalf is fixed in 10% neutral buffered formalin for histologicalassessment of liver fibrosis using light microscopy (10× magnification).Liver tissue homogenates are analyzed for collagen levels using Sircol(Biocolor Ltd, UK). Fixed Liver tissue is stained using hematoxylin andeosin (H&E) and trichrome and liver fibrosis is determined byquantitative, computer-assisted densitometry of collagen in liver tissuesections using light microscopy. Plasma and liver tissue lysates arealso analyzed for concentrations of inflammatory, pro-fibrotic andtissue injury biomarkers including transforming growth factor β1,hyaluronic acid, tissue inhibitor of metalloproteinase-1, matrixmatelloproteinase-7, connective tissue growth factor and lactatedehydrogenase activity, using commercially available ELISA. Theresulting data are plotted using Graphpad prism and statisticaldifferences between groups determined.

Example 18 Mouse Intravenous LPA-Induced Histamine Release

A mouse intravenous LPA-induced histamine release model is utilized todetermine the in vivo potency of LPA₁ and LPA₃ receptor antagonists.Female CD-1 mice (weighing 25-35 grams) are administered compound (i.p.,s.c. or p.o.) in a volume of 10 ml/kg 30 minutes to 24 hours prior tointravenous LPA challenge (300 μg/mouse in 0.1% FAF BSA). Immediatelyfollowing LPA challenge mice are placed into an enclosed Plexiglaschamber and exposed to an isoflurane for a period of 2 minutes. They areremoved, decapitated and trunk blood collected into tubes containingEDTA. Blood is then centrifuged at 10,000×g for 10 minutes at 4° C.Histamine concentrations in the plasma are determined by EIA. Drugconcentrations in plasma are determined by mass spectrometry. The doseto achieve 50% inhibition of blood histamine release is calculated bynonlinear regression (Graphpad Prism) and plotted as the ED₅₀. Theplasma concentration associated with this dose is plotted as the EC₅₀.

Example 19 Mouse Unilateral Ureteral Obstruction Kidney Fibrosis Model

Female C57BL/6 mice (Harlan, 20-25 g) housed 4/cage will be given freeaccess to food and water and allowed to acclimate for at least 7 daysprior to test initiation. After the habituation phase, mice undergounilateral ureteral obstruction (UUO) surgery or sham to left kidney.Briefly, a longitudinal, upper left incision is performed to expose theleft kidney. The renal artery is located and 6/0 silk thread is passedbetween the artery and the ureter. The thread is looped around theureter and knotted 3 times insuring full ligation of ureter. The kidneyis returned to abdomen, the abdominal muscle is sutured and the skin isstapled closed. Mice are returned to their cages and monitored daily forthe duration of the experiment. Test compound or vehicle is deliveredpo, ip or sc daily. The route and frequency of dosing is based onpreviously determined pharmacokinetic properties. All animals aresacrificed using inhaled isoflurane 4, 8 or 14 days after UUO surgery.Following sacrifice blood is drawn via cardiac puncture, the kidneys areharvested and one half of the kidney is frozen at −80° C. and the otherhalf is fixed in 10% neutral buffered formalin for histologicalassessment of kidney fibrosis using light microscopy (10×magnification). Kidney tissue homogenates are analyzed for collagenlevels using Sircol (Biocolor Ltd, UK). Fixed kidney tissue is alsostained using hematoxylin and eosin (H&E) and trichrome and kidneyfibrosis is determined by quantitative, computer-assisted densitometryof collagen in liver tissue sections using light microscopy. Plasma andkidney tissue lysates are also analyzed for concentrations ofinflammatory, pro-fibrotic and tissue injury biomarkers includingtransforming growth factor β1, hyaluronic acid, tissue inhibitor ofmetalloproteinase-1, matrix matelloproteinase-7, connective tissuegrowth factor and lactate dehydrogenase activity, using commerciallyavailable ELISA. The resulting data are plotted using Graphpad prism andstatistical differences between groups determined.

Example 20 Clinical Trial in Humans with Idiopathic Pulmonary Fibrosis(IPF) Purpose

The purposes of this study is to assess the efficacy of treatment with acompound of Formula (I), Formula (II) or Formula (III) compared withplacebo in patients with idiopathic pulmonary fibrosis (IPF) and toassess the safety of treatment with a compound of Formula (I), Formula(II) or Formula (III) compared with placebo in patients with IPF.

The primary outcome variable is the absolute change in percent predictedforced vital capacity (FVC) from baseline to Week 72.

Secondary outcome measures include: composite outcomes of importantIPF-related events; progression-free survival; categorical assessment ofabsolute change in percent predicted FVC from baseline to Week 72;change in Shortness-of-Breath from baseline to Week 72; change inpercent predicted hemoglobin (Hb)-corrected carbon monoxide diffusingcapacity (DLco) of the lungs from baseline to Week 72; change in oxygensaturation during the 6 minute walk test (6MWT) from baseline to Week72; change in high-resolution computed tomography (HRCT) assessment frombaseline to Week 72; change in distance walked in the 6MWT from baselineto Week 72.

Criteria

Patients eligible for this study include those patients that satisfy thefollowing inclusion criteria: diagnosis of IPF; 40 to 80 years of age;FVC≧50% predicted value; DLco≧35% predicted value; either FVC orDLco≦90% predicted value; no improvement in past year; able to walk 150meters in 6 minutes and maintain saturation ≧83% while on no more than 6L/min supplemental oxygen.

Patients are excluded from this study if they satisfy any of thefollowing criteria: unable to undergo pulmonary function testing;evidence of significant obstructive lung disease or airwayhyper-responsiveness; in the clinical opinion of the investigator, thepatient is expected to need and be eligible for a lung transplant within72 weeks of randomization; active infection; liver disease; cancer orother medical condition likely to result in death within 2 years;diabetes; pregnancy or lactation; substance abuse; personal or familyhistory of long QT syndrome; other IPF treatment; unable to take studymedication; withdrawal from other IPF trials.

Patients are orally dosed with either placebo or an amount of compoundof Formula (I), Formula (II) or Formula (III) (1 mg/day-1000 mg/day).The primary outcome variable will be the absolute change in percentpredicted FVC from Baseline to Week 72. Patients will receive blindedstudy treatment from the time of randomization until the last patientrandomized has been treated for 72 weeks. A Data Monitoring Committee(DMC) will periodically review safety and efficacy data to ensurepatient safety.

After week 72, patients who meet the Progression of Disease (POD)definition, which is a ≧10% absolute decrease in percent predicted FVCor a ≧15% absolute decrease in percent predicted DLco, will be eligibleto receive permitted IPF therapies in addition to their blinded studydrug. Permitted IPF therapies include corticosteroids, azathioprine,cyclophosphamide and N-acetyl-cysteine.

Pharmaceutical Compositions

Example 21 Parenteral Composition

To prepare a parenteral pharmaceutical composition suitable foradministration by injection (subcutaneous, intravenous, and the like),100 mg of a water-soluble salt of a compound of Formula (I), Formula(II) or Formula (III) is dissolved in sterile water and then mixed with10 mL of 0.9% sterile saline. The mixture is incorporated into a dosageunit form suitable for administration by injection

In another embodiment, the following ingredients are mixed to form aninjectable formulation: 1.2 g of a compound of Formulas (I), 2.0 mL ofsodium acetate buffer solution (0.4 M), HCl (1 N) or NaOH (1 M) (q.s. tosuitable pH), water (distilled, sterile) (q.s. to 20 mL). All of theabove ingredients, except water, are combined and stirred and ifnecessary, with slight heating if necessary. A sufficient quantity ofwater is then added.

Example 22 Oral Composition

To prepare a pharmaceutical composition for oral delivery, 100 mg of acompound of Formula (I), Formula (II) or Formula (III) is mixed with 750mg of starch. The mixture is incorporated into an oral dosage unit for,such as a hard gelatin capsule, which is suitable for oraladministration.

Example 23 Sublingual (Hard Lozenge) Composition

To prepare a pharmaceutical composition for buccal delivery, such as ahard lozenge, mix 100 mg of a compound of Formula (I), Formula (II) orFormula (III) with 420 mg of powdered sugar mixed, with 1.6 mL of lightcorn syrup, 2.4 mL distilled water, and 0.42 mL mint extract. Themixture is gently blended and poured into a mold to form a lozengesuitable for buccal administration.

Example 24 Fast-Disintegrating Sublingual Tablet

A fast-disintegrating sublingual tablet is prepared by mixing 48.5% byweigh of a compound of Formula (I), Formula (II) or Formula (III), 44.5%by weight of microcrystalline cellulose (KG-802), 5% by weight oflow-substituted hydroxypropyl cellulose (50 μm), and 2% by weight ofmagnesium stearate. Tablets are prepared by direct compression (AAPSPharm Sci Tech. 2006; 7(2):E41). The total weight of the compressedtablets is maintained at 150 mg. The formulation is prepared by mixingthe amount of compound of Formula (I), Formula (II) or Formula (III)with the total quantity of microcrystalline cellulose (MCC) andtwo-thirds of the quantity of low-substituted hydroxypropyl cellulose(L-HPC) by using a three dimensional manual mixer (Inversina®,Bioengineering AG, Switzerland) for 4.5 minutes. All of the magnesiumstearate (MS) and the remaining one-third of the quantity of L-HPC areadded 30 seconds before the end of mixing.

Example 25 Inhalation Composition

To prepare a pharmaceutical composition for inhalation delivery, 20 mgof a compound of Formula (I), Formula (II) or Formula (III) is mixedwith 50 mg of anhydrous citric acid and 100 mL of 0.9% sodium chloridesolution. The mixture is incorporated into an inhalation delivery unit,such as a nebulizer, which is suitable for inhalation administration.

Example 26 Rectal Gel Composition

To prepare a pharmaceutical composition for rectal delivery, 100 mg of acompound of Formula (I), Formula (II) or Formula (III) is mixed with 2.5g of methylcelluose (1500 mPa), 100 mg of methylparapen, 5 g of glycerinand 100 mL of purified water. The resulting gel mixture is thenincorporated into rectal delivery units, such as syringes, which aresuitable for rectal administration.

Example 27 Topical Gel Composition

To prepare a pharmaceutical topical gel composition, 100 mg of acompound of Formula (I), Formula (II) or Formula (III) is mixed with1.75 g of hydroxypropyl celluose, 10 mL of propylene glycol, 10 mL ofisopropyl myristate and 100 mL of purified alcohol USP. The resultinggel mixture is then incorporated into containers, such as tubes, whichare suitable for topical administration.

Example 28 Ophthalmic Solution Composition

To prepare a pharmaceutical opthalmic solution composition, 100 mg of acompound of Formula (I), Formula (II) or Formula (III) is mixed with 0.9g of NaCl in 100 mL of purified water and filtered using a 0.2 micronfilter. The resulting isotonic solution is then incorporated intoophthalmic delivery units, such as eye drop containers, which aresuitable for ophthalmic administration.

Example 29 Nasal Spray Solution

To prepare a pharmaceutical nasal spray solution, 10 g of a compound ofFormula (I), Formula (II) or Formula (III) is mixed with 30 mL of a0.05M phosphate buffer solution (pH 4.4). The solution is placed in anasal administrator designed to deliver 100 μl of spray for eachapplication.

The examples and embodiments described herein are for illustrativepurposes only and various modifications or changes suggested to personsskilled in the art are to be included within the spirit and purview ofthis application and scope of the appended claims.

What is claimed is:
 1. A compound having the structure of Formula (I), or a pharmaceutically acceptable salt:

wherein, R¹ is —CO₂H, —CO₂R^(A), —C(═O)NHSO₂R¹⁰, —C(═O)N(R⁹)₂, —C(═O)NH—OH, —C(═O)NH—CN, tetrazolyl, —OH, —OR⁹, substituted or unsubstituted monocyclic heteroaryl, or substituted or unsubstituted monocyclic heterocycloalkyl; R^(A) is H or C₁-C₆alkyl; L¹ is absent, a substituted or unsubstituted C₁-C₆alkylene, a substituted or unsubstituted C₁-C₆-fluoroalkylene, a substituted or unsubstituted C₁-C₆heteroalkylene, a substituted or unsubstituted C₃-C₆cycloalkylene, a substituted or unsubstituted C₁-C₆heterocycloalkylene, a substituted or unsubstituted phenylene, a substituted or unsubstituted heteroarylene, or -L²-B-L³-; L² is absent, a substituted or unsubstituted C₁-C₆alkylene, a substituted or unsubstituted C₁-C₆-fluoroalkylene, or a substituted or unsubstituted C₁-C₆heteroalkylene; B is a substituted or unsubstituted C₃-C₆cycloalkylene, a substituted or unsubstituted C₂-C₆heterocycloalkylene, a substituted or unsubstituted phenylene, or a substituted or unsubstituted heteroarylene; L³ is absent, a substituted or unsubstituted C₁-C₆alkylene, a substituted or unsubstituted C₁-C₆-fluoroalkylene, or a substituted or unsubstituted C₁-C₆heteroalkylene; R³ is H, C₁-C₄alkyl, or C₁-C₄-fluoroalkyl; R⁴ is —NR⁷C(═O)OCH(R⁸)—CY, or —NR⁷C(═O)O—CY; R⁷ is H or C₁-C₄alkyl; R⁸ is H, C₁-C₄alkyl, or C₁-C₄-fluoroalkyl; CY is a substituted or unsubstituted C₃-C₁₀cycloalkyl, a substituted or unsubstituted C₂-C₁₀heterocycloalkyl, a substituted or unsubstituted phenyl, a substituted or unsubstituted napthyl, or a substituted or unsubstituted monocyclic or bicyclic heteroaryl; or R⁸ and CY are taken together with the carbon atom to which they are attached to form a substituted or unsubstituted C₃-C₁₀-carbocycle or a substituted or unsubstituted C₂-C₁₀heterocycle; R⁵ and R⁶ are each independently selected from H, halogen, —CN, —NO₂, —OH, —OR¹⁰, —S(═O)₂R¹⁰, —N(R⁹)S(═O)₂R¹⁰, —S(═O)₂N(R⁹)₂, —C(═O)R¹⁰, —OC(═O)R¹⁰, —CO₂R⁹, —OCO₂R¹⁰, —N(R⁹)₂, —C(═O)N(R⁹)₂, —C(═O)N(R⁹)₂, —NR⁹C(═O)N(R⁹)₂, —NR⁹C(═O)R¹⁰, —NR⁹C(═O)OR¹⁰, C₁-C₄alkyl, C₁-C₄-fluoroalkyl, C₁-C₄-fluoroalkoxy, C₁-C₄alkoxy, and C₁-C₄heteroalkyl; each R⁹ is independently selected from H, C₁-C₆alkyl, C₁-C₆heteroalkyl, C₁-C₆-fluoroalkyl, a substituted or unsubstituted C₃-C₁₀cycloalkyl, a substituted or unsubstituted C₂-C₁₀heterocycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, substituted or unsubstituted —C₁-C₄alkylene-(substituted or unsubstituted C₃-C₁₀cycloalkyl), —C₁-C₄alkylene-(substituted or unsubstituted C₂-C₁₀heterocycloalkyl), —C₁-C₄alkylene-(substituted or unsubstituted aryl), or —C₁-C₄alkylene-(substituted or unsubstituted heteroaryl); or two R⁹ groups attached to the same N atom are taken together with the N atom to which they are attached to form a substituted or unsubstituted heterocycle; and R¹⁰ is selected from C₁-C₆alkyl, C₁-C₆heteroalkyl, C₁-C₆-fluoroalkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, —C₁-C₄alkylene-(substituted or unsubstituted C₃-C₁₀cycloalkyl), —C₁-C₄alkylene-(substituted or unsubstituted C₂-C₁₀heterocycloalkyl), —C₁-C₄alkylene-(substituted or unsubstituted aryl), and —C₁-C₄alkylene-(substituted or unsubstituted heteroaryl).
 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: R¹ is —CO₂H, —CO₂R^(A), or a substituted or unsubstituted monocyclic heteroaryl; R³ is C₁-C₄alkyl; R⁴ is —NR⁷C(═O)OCH(R⁸)—CY; R⁷ is H; R^(A) is H, —CH₃, or —CH₂CH₃.
 3. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein: R⁵ and R⁶ are each independently selected from H, halogen, —OH, —CF₃, —OCF₃, and —OCH₃; and CY is a substituted or unsubstituted C₃-C₆cycloalkyl, a substituted or unsubstituted phenyl, or a substituted or unsubstituted monocyclic heteroaryl containing 0-4 N atoms.
 4. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein: R³ is —CH₃ or —CH₂CH₃; and L¹ is absent, C₁-C₆alkylene, C₁-C₆-fluoroalkylene, C₁-C₆heteroalkylene, a substituted or unsubstituted C₃-C₆cycloalkylene, a substituted or unsubstituted phenylene, a substituted or unsubstituted monocyclic heteroarylene containing 0-4 N atoms, or -L²-B-L³-.
 5. The compound of claim 4, or a pharmaceutically acceptable salt thereof, wherein: L² is absent, C₁-C₆alkylene, C₁-C₆-fluoroalkylene, or C₁-C₆heteroalkylene; B is a substituted or unsubstituted C₃-C₆cycloalkylene, a substituted or unsubstituted C₂-C₆heterocycloalkylene, a substituted or unsubstituted phenylene, or a substituted or unsubstituted monocyclic heteroarylene; and L³ is absent, C₁-C₆alkylene, C₁-C₆-fluoroalkylene, or C₁-C₆heteroalkylene.
 6. The compound of claim 5, or a pharmaceutically acceptable salt thereof, wherein: B is a substituted or unsubstituted phenylene, or a substituted or unsubstituted monocyclic heteroarylene containing 0-4 N atoms.
 7. The compound of claim 6, or a pharmaceutically acceptable salt thereof, wherein: L¹ is -L²-B-L³-; R⁴ is

R⁸ is —CH₃ or —CF₃; CY is a substituted or unsubstituted phenyl.
 8. The compound of claim 7, or a pharmaceutically acceptable salt thereof, wherein: R¹ is —CO₂H, —CO₂R^(A), or a substituted or unsubstituted monocyclic heteroaryl containing 1-4 N atoms; L² is absent , C₁-C₆alkylene, C₁-C₆-fluoroalkylene, —C₁-C₆alkylene-S(═O)—, —C₁-C₆alkylene-SO₂—, or —C₁-C₄alkylene-NR²—; B is a substituted or unsubstituted phenylene, or a substituted or unsubstituted monocyclic heteroarylene containing 1-4 N atoms; L² is absent, C₁-C₆alkylene, C₁-C₆-fluoroalkylene, —S—C₁-C₆alkylene-, —S(═O)—C₁-C₆alkylene-, —SO₂—C₁-C₆alkylene-, or —NR²—C₁-C₄alkylene; R² is H, C₁-C₂alkyl, or C₁-C₂-fluoroalkyl.
 9. The compound of claim 4, or a pharmaceutically acceptable salt thereof, wherein: L¹ is a C₁-C₆alkylene, C₁-C₆-fluoroalkylene, or C₁-C₆heteroalkylene; R⁴ is

R⁸ is —CH₃ or —CF₃; CY is a substituted or unsubstituted phenyl.
 10. The compound of claim 9, or a pharmaceutically acceptable salt thereof, wherein: R¹ is —CO₂H, —CO₂R^(A), or a substituted or unsubstituted monocyclic heteroaryl containing 1-4 N atoms.
 11. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) has the structure of Formula (II) or Formula (III):


12. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable inactive ingredient.
 13. The pharmaceutical composition of claim 12, wherein the pharmaceutical composition is: (a) formulated for intravenous injection, subcutaneous injection, oral administration, inhalation, nasal administration, topical administration, ophthalmic administration or otic administration; or (b) a tablet, a pill, a capsule, a liquid, an inhalant, a nasal spray solution, a suppository, a suspension, a gel, a colloid, a dispersion, a suspension, a solution, an emulsion, an ointment, a lotion, an eye drop or an ear drop.
 14. The pharmaceutical composition of claim 12 further comprising one or more therapeutically active agents selected from: corticosteroids, immunosuppressants, analgesics, anti-cancer agents, anti-inflammatories, chemokine receptor antagonists, bronchodilators, leukotriene receptor antagonists, leukotriene formation inhibitors, monoacylglycerol kinase inhibitors, phospholipase A₁ inhibitors, phospholipase A₂ inhibitors, lysophospholipase D (lysoPLD) inhibitors, autotaxin inhibitors, decongestants, antihistamines, mucolytics, anticholinergics, antitussives, expectorants, and β-2 agonists.
 15. A method for alleviating, abating or ameliorating at least one symptom of organ fibrosis in a mammal comprising administering a therapeutically effective amount of a compound according to claim 1, or a pharmaceutically acceptable salt thereof to a mammal in need thereof.
 16. A method of improving lung function in a mammal comprising administering a therapeutically effective amount of a compound according to claim 1, or a pharmaceutically acceptable salt thereof to the mammal in need thereof.
 17. A method for alleviating, abating or ameliorating at least one symptom of cancer in which the inhibition of the physiological activity of LPA is useful, in a mammal comprising administering a therapeutically effective amount of a compound according to claim 1, or a pharmaceutically acceptable salt thereof to a mammal in need thereof. 